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Herd health management is a planned program to optimize health, welfare, and production of dairy cows. A cross-sectional study focused on herd health management of dairy farms, was carried out to assess the biosecurity, health, and disease management practices with associated socio-demographic factors. A structured questionnaire with closed- and open-ended questions was employed for farmers' interviews to acquire information on dairy farms. Data were collected from randomly selected 205 dairy farms in five districts (Dhaka-Savar, Mymensingh, Gazipur, Rangpur, and Sirajganj) of Bangladesh. Overall, 1.5% farms had "good", 17% had "moderate" and 81.5% had "poor" levels of biosecurity, health and disease management practices (BHDMP). Analysis of variance revealed that mean BHDMP scores were significantly (p < 0.05) higher in farms of Rangpur (27.6 ± 0.8) and Gazipur (26.7 ± 0.6) than those of the other three districts. Educational status had a significant effect on BHDMP scores. The farms owned by graduated farmers had the highest mean BHDMP score (30.4 ± 1.1) while the farms operated by illiterate farmers had the lowest BHDMP score (20.4 ± 0.4). Multiple linear regression analysis also revealed that educational status (secondary to post-graduation), training experience, area of study (Rangpur district), and herd size were significantly associated with BHDMP score of the farms. The findings lay a basis for improving dairy farms' biosecurity, health and disease management practices, which further envisage the adoption and implementation of herd health management programs., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Magnolia champaca Linn. has traditionally been used for medicinal activity in Asia for treating various chronic diseases as well as a source of food, medicines, and other commodities. Due to the long-used history of this plant, the present study was designed to explore the in vitro , in vivo and in silico anti-inflammatory and antineoplastic properties of the methanolic extract and fractions and the pure compound isolated from the most active chloroform fraction (CHF) of the stem bark of the plant. The isolated compound from the most active CHF was characterized and identified as a glycoside, trans-syringin, through chromatographic and spectroscopic ( 1 H-NMR and 13 C-NMR) analyses. In the in vitro anti-inflammatory assay, CHF was most effective in inhibiting inflammation and hemolysis of RBCs by 73.91 ± 1.70% and 75.92 ± 0.14%, respectively, induced by heat and hypotonicity compared to standard acetylsalicylic acid. In the egg albumin denaturation assay, CME and CHF showed the highest inhibition by 56.25 ± 0.82% and 65.82 ± 3.52%, respectively, contrasted with acetylsalicylic acid by 80.14 ± 2.44%. In an in vivo anti-inflammatory assay, statistically significant (p < 0.05) decreases in the parameters of inflammation, such as paw edema, leukocyte migration and vascular permeability, were recorded in a dose-dependent manner in the treated groups. In the antineoplastic assay, 45.26 ± 2.24% and 68.31 ± 3.26% inhibition of tumor cell growth for pure compound were observed compared to 73.26 ± 3.41% for standard vincristine. Apoptotic morphologic alterations, such as membrane and nuclear condensation and fragmentation, were also found in EAC cells after treatment with the isolated bioactive pure compound. Such treatment also reversed the increased WBC count and decreased RBC count to normal values compared to the untreated EAC cell-bearing mice and the standard vincristine-treated mice. Subsequently, in silico molecular docking studies substantiated the current findings, and the isolated pure compound and standard vincristine exhibited -6.4 kcal/mol and -7.3 kcal/mol binding affinities with topoisomerase-II. Additionally, isolated pure compound and standard diclofenac showed -8.2 kcal/mol and -7.6 kcal/mol binding affinities with the COX-2 enzyme, respectively. The analysis of this research suggests that the isolated bioactive pure compound possesses moderate to potent anti-inflammatory and antineoplastic activity and justifies the traditional uses of the stem bark of M. champaca . However, further investigations are necessary to analyze its bioactivity, proper mechanism of action and clinical trials for the revelation of new drug formulations., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Mst. Shahnaj Parvin reports administrative support was provided by 10.13039/501100016173University of Rajshahi. Mst. Shahnaj Parvin reports a relationship with University of Rajshahi that includes: employment. Mst. Shahnaj Parvin has patent pending to Not applicable. Not applicable If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)

The process of platelet aggregation is often influenced by several factors including sex and age. A literature review confirmed the existence of sex-related differences in platelet aggregation. Although 68 out of 78 papers found such differences, there are still some controversies regarding these differences, which can be due to multiple factors (age, trigger, concomitant disease, sample handling, etc.). These outcomes are discussed in line with novel results obtained from a local study, in which blood samples from a total of 53 overall healthy women and men with ages ranging from 20 to 66 years were collected. Aggregation was induced with seven different triggers (ristocetin, thrombin receptor activating peptide 6 [TRAP-6], arachidonic acid [AA], platelet-activating factor 16 [PAF-16], ADP, collagen, or thromboxane A 2 analog U-46619) ex vivo. In addition, three FDA-approved antiplatelet drugs (vorapaxar, ticagrelor, or acetylsalicylic acid [ASA]) were also tested. In general, women had higher aggregation responses to some agonists (ADP, TRAP), as well as lower benefit from inhibitors (ASA, vorapaxar). The aggregatory responses to AA and TRAP decreased with age in both sexes, while responses to ADP, U-46619, and PAF were affected by age only in women. In conclusion, more studies are needed to decipher the biological importance of sex-related differences in platelet aggregation in part to enable personalized antiplatelet treatment., Competing Interests: None declared., (Thieme. All rights reserved.)

The study was conducted to identify the antimicrobial resistance genes (ARGs) in Lactobacillus spp. from poultry probiotic products and their potential to spread among Escherichia coli. Lactobacillus spp. were isolated and identified from 35 poultry probiotic samples based on the cultural, biochemical, and molecular findings. All the isolates ( n  = 35) were screened for the presence of some ARGs such as β-lactamases encoding genes ( bla TEM, bla CTXM-1, and bla CTXM-2), plasmid-mediated quinolone resistance gene ( qnr A, qnr B, and qnr S), and tetracycline resistance genes ( tet A and tet B). Five Lactobacillus spp. isolates from three brands were positive for one or more ARGs. The qnr S was detected in four isolates. The bla TEM and tet B were detected in two isolates. One isolate contained bla CTX-M-1, bla CTX-M-2, and tet A genes. Brand-wise analysis revealed that one isolate from Brand 4 contained bla TEM, bla CTX-M-1, bla CTX-M-2, qnr S, and tet A genes, one isolate from Brand 2 contained bla TEM gene, and three isolates from Brand 7 harbored qnr S gene. The co-culture of Lactobacillus spp. and E. coli resulted in the transmission of qnr S, CTX-M-1, and tet A from Lactobacillus spp. to E. coli . Results of antimicrobial susceptibility test revealed that the highest resistance was observed to cefepime and cefotaxime followed by penicillin G, oxacillin, cefuroxime, and ofloxacin. The findings of the present study indicate the potential risk of horizontal spread of antimicrobial resistance through probiotic bacteria among the poultry population. Therefore, it is very necessary to check for ARGs along with other attributes of probiotic bacteria to avoid the inclusion of resistant strains in probiotics., Competing Interests: The authors declare that they have no known competing interests., (© 2023 The Author(s).)

Objectives: This study was conducted to assess poultry farmers' knowledge and practices regarding antibiotics, antimicrobial usage (AMU), and antimicrobial resistance (AMR), and to identify the sociodemographic factors of inappropriate use of antibiotics in commercial poultry farms in Bangladesh., Methods: A qualitative survey of 140 farmers in Bangladesh was conducted from March to May 2019. A logistic regression model was used to identify factors associated with the inappropriate use of antibiotics., Results: 47.1% of farmers were unable to explain antibiotics, 42.9% used antibiotics for preventive purposes, 4.3% used them as growth promoters, 25.7% used them as suggested by veterinarians, 42.9% used leftover antibiotics, 50% did not maintain antibiotics residual withdrawal period, and 98.6% did not know about AMR. In bivariable regression analysis, sex and primary occupation of poultry farmers, their knowledge about withdrawal periods for antibiotics, and no contact with veterinary surgeons (VS) were found to be significantly associated with the inappropriate use of antibiotics, while only 'no contact with VS' was identified in multivariable regression analysis., Conclusions: The findings suggest an urgent need to improve understanding of antibiotics and AMR. Adequate supervision by veterinarians would ensure adherence to appropriate AMU patterns, and would limit the misuse of antibiotics and associated AMR development in farms., Competing Interests: The authors declare no conflicts of interest., (© 2023 The Author(s).)

A polyphenol-rich diet has beneficial effects on cardiovascular health. However, dietary polyphenols generally have low bioavailability and reach low plasma concentrations. Small phenolic metabolites of these compounds formed by human microbiota are much more easily absorbable and could be responsible for this effect. One of these metabolites, 4-methylcatechol (4-MC), was suggested to be a potent anti-platelet compound. The effect of 4-MC was tested ex vivo in a group of 53 generally healthy donors using impedance blood aggregometry. The mechanism of action of this compound was also investigated by employing various aggregation inducers/inhibitors and a combination of aggregometry and enzyme linked immunosorbent assay (ELISA) methods. 4-MC was confirmed to be more potent than acetylsalicylic acid on both arachidonic acid and collagen-triggered platelet aggregation. Its clinically relevant effect was found even at a concentration of 10 μM. Mechanistic studies showed that 4-MC is able to block platelet aggregation caused by the stimulation of different pathways (receptors for the von Willebrand factor and platelet-activating factor, glycoprotein IIb/IIIa, protein kinase C, intracellular calcium elevation). The major mechanism was defined as interference with cyclooxygenase-thromboxane synthase coupling. This study confirmed the strong antiplatelet potential of 4-MC in a group of healthy donors and defined its mechanism of action.

Lumpy skin disease (LSD) is an emerging viral disease of cattle that negatively impacts livestock by reducing animals' production and increasing management costs. Like other countries, Bangladesh has also faced the ominous effects of LSD. Because of this pressing concern, the present study was undertaken to determine the attack rate and risk factors along with economic losses of the recent LSD outbreaks from October 2019 to June 2020 in Bangladesh's Mymensingh and Gaibandha districts. A total of 1187 cattle from 150 randomly selected cattle farms were surveyed. The diagnosis was made both clinically and molecularly. The overall attack rate, mortality and case fatality were 34%, 1.10% and 3.23%, respectively, but in calves up to 1 month of age, the attack rate, mortality and case-fatality were 41.6%, 12.5% and 30%, respectively. In risk factor analysis using multiple logistic regression, the following factors were significantly (p < .05) associated with LSD occurrence: no administration of goat pox vaccine, frequency of goat pox vaccination - single dose, introduction of new cattle, land type (low land and medium-high land), use of common syringe and needle, and use of common utensils. The average economic loss per case was 9384.41 BDT (110.40 US $). The higher economic loss per case was accounted in Mymensingh district (10248.91 BDT ≈ 120.58 US $) than Gaibandha district (8211.52 BDT ≈ 96.61 US $). The loss was higher in crossbred cattle (9709.58 BDT ≈ 114.23 US $) compared to indigenous cattle (7595.94 BDT ≈ 89.36 US $). The total estimated annual loss due to LSD in Mymensingh and Gaibandha districts was 7763.25 million BDT (91.33 million US $). In Gaibandha district, the estimated annual loss was 2666.20 million BDT (31.37 million US $) while in Mymensingh district it was 5097.05 million BDT (59.97 million US $). Findings of the study could provide useful epidemiological data with losses incurred due to LSD to the livestock owners, field veterinarians and government-level livestock regulators, which will help to formulate prevention and control strategies and minimize the negative impact of LSD's probable return or spread., (© 2022 Wiley-VCH GmbH.)

Data on alkaloid interactions with the physiologically important transition metals, iron and copper, are mostly lacking in the literature. However, these interactions can have important consequences in the treatment of both Alzheimer's disease and cancer. As isoquinoline alkaloids include galanthamine, an approved drug for Alzheimer's disease, as well as some potentially useful compounds with cytostatic potential, 28 members from this category of alkaloids were selected for a complex screening of interactions with iron and copper at four pathophysiologically relevant pH and in non-buffered conditions (dimethyl sulfoxide) by spectrophotometric methods in vitro. With the exception of the salts, all the alkaloids were able to chelate ferrous and ferric ions in non-buffered conditions, but only five of them (galanthine, glaucine, corydine, corydaline and tetrahydropalmatine) evoked some significant chelation at pH 7.5 and only the first two were also active at pH 6.8. By contrast, none of the tested alkaloids chelated cuprous or cupric ions. All the alkaloids, with the exception of the protopines, significantly reduced the ferric and cupric ions, with stronger effects on the latter. These effects were mostly dependent on the number of free aromatic hydroxyls, but not other hydroxyl groups. The most potent reductant was boldine. As most of the alkaloids chelated and reduced the ferric ions, additional experimental studies are needed to elucidate the biological relevance of these results, as chelation is expected to block reactive oxygen species formation, while reduction could have the opposite effect.

Oral and periodontal diseases (OPD) is considered one of the main problems of dentistry worldwide. This study aimed to estimate the prevalence of oral and periodontal pathogenic bacteria along with their antimicrobial resistance pattern in 131 children patients aged between 4-10 years who attended in Mymensingh Medical College Hospital during October 2019 to March 2020. OPD pathogens were identified through isolation, cultural and biochemical properties, and nucleic acid detection. The isolates were subjected to antimicrobial susceptibility to 12 antibiotics commonly used in dentistry. In addition, the isolates were analyzed molecularly for the presence of six virulence and three antibacterial resistance genes. Five pathogens were identified, of which Staphylococcus aureus ( S. aureus ) (49%) and S. salivarius (46%) were noticed frequently; other bacteria included S. mutans (16.8%), S. sobrinus (0.8%) and L. fermentum (13.7%). The virulence genes-clumping factor A ( clfA ) was detected in 62.5% isolates of S. aureus , and gelatinase enzyme E ( gelE ) gene was detected in 5% isolates of S. salivarius, while other virulence genes were not detected. All the tested isolates were multidrug-resistant. The overall prevalence of MDR S. aureus , Streptococcus spp. and L. fermentum was 92.2%, 95.1% and 100%, respectively. It was observed that a high proportion of isolates were found resistant to 5-8 antibiotics. A majority of S. aureus , Streptococcus spp., and L. fermentum isolates tested positive for the β-lactamase resistance genes blaTEM and cfxA , as well as the methicillin resistance gene mecA . Phylogenetically, the resistance genes showed variable genetic character among Bangladeshi bacterial pathogens. In conclusion, S. aureus and S. salivarius were major OPD pathogens in patients attended in Mymensingh Medical College Hospital of Bangladesh, and most were Beta-lactam and methicillin resistant.

Background: Children suffer from various oral and periodontal diseases. Dental caries is one of the most prevalent oral diseases among children in the world. This study was conducted to identify the prevalence and risk factors of dental caries in children in Mymensingh, Bangladesh., Methods: A cross-sectional study was conducted on 362 pediatric patients who attended the Dental Unit of Mymensingh Medical College from March to September 2019. The sample size was calculated using a statistical formula and the children were selected using a systematic random sampling technique. Children and their guardians were interviewed and data were recorded using a structured questionnaire. Risk factors were analyzed using multivariate logistic regression., Results: The overall prevalence of dental caries was 82.7%. The prevalence of caries was significantly higher in aged children (8-10 years) and also in rural, low-income, and illiterate families. Seven significant risk factors were identified that included residence in the rural area (OR: 7.31 [1.73-30.83]), a parental income of BDT ≤ 20,000 per month (OR: 4.75 [1.49-15.05]), reduced duration (≤1 min) of teeth cleaning (OR: 18.54 [2.05-168.17]), teeth cleaning before breakfast (OR: 93.30 [10.95-795.32]), the spoon-feeding method (OR: 12.57 [2.09-75.61]), long-term (37-48 months) breastfeeding (OR: 212.53 [8.69-5195.25]), and family oral problem (OR: 8.20 [2.57-26.16])., Conclusions: The prevalence of dental caries among the children in Mymensingh is very high and was associated with residence in rural areas, parental income, reduced duration of teeth cleaning, teeth cleaning before breakfast, the spoon-feeding method, long-term breastfeeding, and family oral problems.

Isoquinoline alkaloids have multiple biological activities, which might be associated with positive pharmacological effects as well as negative adverse reactions. As bleeding was suggested to be a side effect of the isoquinoline alkaloid berberine, we decided to ascertain if different isoquinoline alkaloids could influence hemocoagulation through the inhibition of either platelet aggregation or blood coagulation. Initially, a total of 14 compounds were screened for antiplatelet activity in whole human blood by impedance aggregometry. Eight of them demonstrated an antiplatelet effect against arachidonic acid-induced aggregation. Papaverine and bulbocapnine were the most potent compounds with biologically relevant IC 50 values of 26.9 ± 12.2 μM and 30.7 ± 5.4 μM, respectively. Further testing with the same approach confirmed their antiplatelet effects by employing the most physiologically relevant inducer of platelet aggregation, collagen, and demonstrated that bulbocapnine acted at the level of thromboxane receptors. None of the alkaloids tested had an effect on blood coagulation measured by a mechanical coagulometer. In conclusion, the observed antiplatelet effects of isoquinoline alkaloids were found mostly at quite high concentrations, which means that their clinical impact is most likely low. Bulbocapnine was an exception. It proved to be a promising antiplatelet molecule, which may have biologically relevant effects.

Objective: Periparturient period is associated with multiple changes including serum concentration of macro minerals and drop in feed intake. Therefore, it is essential to know the actual concentrations of major macro minerals, glucose and ketone bodies in blood during the periparturient period. The objectives of the study were to study the dynamics of calcium, magnesium, phosphorus, and glucose in serum and ketone bodies in the urine of periparturient cows and to estimate the incidence of subclinical metabolic diseases., Results: Results showed that all the urine samples were negative for ketone bodies. Incidence of subclinical form of hypocalcaemia, hypomagnesaemia, hypophosphatemia, and hypoglycemia in periparturient cows was 31.03%, 48.28%, 17.24% and 55.17%, respectively. Older cows with high BCS and milk yield were mostly affected with a subclinical form of hypocalcaemia and hypoglycemia. No significant differences were observed in serum level of macro minerals and glucose at different time points of periparturient period of cows fed with a balanced ration, and between two groups of cows those were given IV injection of calcium and magnesium on the day of parturition and those were not given. Age, parity, and milk yield had no significant effect on the concentration of calcium, magnesium, phosphorus, and glucose., (© 2022. The Author(s).)

Antimicrobial resistance (AMR) among foodborne bacteria is a well-known public health problem. A sink survey was conducted to determine the AMR pattern of common foodborne bacteria in cloacal swab of broiler chickens and sewage samples from five wholesale chicken markets of Dhaka city in Bangladesh. Bacteria were identified by culture-based and molecular methods, and subjected to antimicrobial susceptibility testing. Resistance genes were identified by multiplex PCR and sequencing. Multidrug resistance (MDR) was observed in 93.2% of E. coli, 100% of Salmonella spp., and 97.2% of S. aureus from cloacal swab samples. For sewage samples, 80% of E. coli, and 100% of Salmonella and S. aureus showed MDR. Noteworthy, 8.3% of S. aureus from cloacal swab samples showed possible extensively drug resistance. Antimicrobial resistance genes (beta-lactamase-blaTEM, blaSHV; quinolone resistance gene-qnrS) were detected in a number of E. coli and Salmonella isolates from cloacal swab and sewage samples. The methicillin resistance gene (mecA) was detected in 47.2% and 25% S. aureus from cloacal swab and sewage samples, respectively. The findings envisage the potential public health risk and environmental health hazard through spillover of common foodborne MDR bacteria., (© 2022. The Author(s).)

Background: Pithecellobium dulce (Roxb.), an evergreen medium-sized, spiny tree which have vast nutritional values and widely used in ayurvedic medicines and home remedies. The plant has also been a rich source of biologically active compounds. The present study was designed to isolate pure compound from ethyl acetate fraction of methanol extract of leaves and to know the efficacy as antioxidant as well as its anti-tumor activity on Ehrlich ascites carcinoma cell (EAC).  METHODS: The leaves were extracted with methanol and fractionated with different solvents. The isolation of the compound was carried out by column chromatography from ethyl acetate fraction (EAF) and structure was revealed by 1 H-NMR and 13 C NMR. The antioxidant activity was investigated by the scavenging of 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radicals as well as the inhibition of oxidative damage of pUC19 plasmid DNA, hemolysis and lipid peroxidation induced by a water-soluble free radical initiator 2,2'-azo (2-asmidinopropane) dihydrochloride (AAPH) in human erythrocytes. In vivo anti-tumor activity of the compound was also evaluated by determining the viable tumor cell count, hematological profiles of experimental mice along with observing morphological changes of EAC cells by fluorescence microscope., Results: The isolated compound kaempferol-3-O-alpha-L-rhamnoside effectively inhibited AAPH induced oxidation in DNA and human erythrocyte model and lipid per oxidation as well as a stronger DPPH radical scavenging activity. In anti-tumor assay, at a dose of 50 mg/kg body weight exhibit about 70.89 ± 6.62% EAC cell growth inhibition, whereas standard anticancer drug vincristine showed 77.84 ± 6.69% growth inhibition., Conclusion: The compound may have a great importance as a therapeutic agent in preventing oxidative damage of biomolecules and therapeutic use in chemotherapy., (© 2022. The Author(s).)

Among the major constituents of Leea rubra (Family Vitaceae) leaves, phenolic and flavonoind compounds are most important for therapeutic purposes and the plant parts have been used in traditional medicine to treat several diseases for long. Thus, in order to scientifically confirm the traditional uses of the L. rubra leaves, the present study was designed to investigate the efficacy of the isolated flavones against AAPH induced oxidative damage to pUC19 DNA by gel electrophoresis and antineoplastic activity was evaluated on Ehrlich ascites carcinoma (EAC) bearing Swiss albino mice by evaluating percentage inhibition of cell growth, morphological changes of EAC cells and hematological parameters of the mice. The isolation was carried out by column chromatography and structure was revealed by 1 H-NMR and 13 C NMR. The result shows that, the isolated compound was identified as myricetin 4'-methoxy-3-O-α-l-rhamnopyranoside based on previously reported data. The isolated flavone effectively inhibited AAPH-induced oxidative damage to DNA; because it could inhibit the formation of circular and linear forms of the DNA. In anti-proliferative assay, 76% growth inhibition of EAC cells was observed as compare to the control mice (p<0.05) at a dose 100 mg/kg body weight. Thus the isolated flavone showed great importance as a possible therapeutic agent in preventing oxidative damage to DNA and the chronic diseases caused by such DNA damage, and can also become important in cancer chemotherapy., Competing Interests: The authors declare that there are no conflicts of interest., (© 2022 The Authors.)

Medicinal plants are sources of antioxidant which may protect the body against oxidative stress related diseases and can be used as human food supplements. In this investigation, seeds of Hygrophila schulli (M. R. Almeida & S. M. Almeida) (Fam.-Acanthaceae), a herbaceous plant well known for its medicinal properties, has been examined for antioxidant activity of crude methanolic extract (CME) and its fraction using in vitro and in vivo assay as well as their protective activity against oxidative damage of DNA and RBC. Total phenolic and flavonoid content have also been estimated using the aluminum chloride colorimetric and Folin-Ciocalteu method. Among the different fractions of CME, Ethyl acetate fraction (EAF) had higher antioxidant activity in vitro assay and was selected for in vivo antioxidant activity in cadmium intoxicated mice. The EAF showed a significant ( p < 0.05 ) increase in serum catalase and SOD activity compared to the control group. TBARS levels were restored to 17.42 and 19.19 nmol/mg protein, respectively, after treatment with EAF and standard ascorbic acid (AA); compared to the normal group (14.96 nmol/mg protein). Similarly, levels of albumin, bilirubin, uric acid, and alkaline phosphatase were also brought back to normal levels. EAF's protective role against oxidative damage of DNA has shown a significant reduction in destroying of nicked DNA. RBC as a target of oxidation by H 2 O 2 and HOCl, EAF showed inhibition of oxidation in a concentration-dependent manner, compared to standard gallic acid. In this study, we confirmed that EAF could scavenge reactive oxygen species (ROS) thus preventing DNA strand scission and the extract can be used as a functional food or nutraceutical product for health benefits., Competing Interests: The authors declare no conflict of interest., (© 2022 The Authors.)

Flavonoids are associated with positive cardiovascular effects. However, due to their low bioavailability, metabolites are likely responsible for these properties. Recently, one of these metabolites, 4-methylcatechol, was described to be a very potent antiplatelet compound. This study aimed to compare its activity with its 22 close derivatives both of natural or synthetic origin in order to elucidate a potential structure-antiplatelet activity relationship. Blood from human volunteers was induced to aggregate by arachidonic acid (AA), collagen or thrombin, and plasma coagulation was also studied. Potential toxicity was tested on human erythrocytes as well as on a cancer cell line. Our results indicated that 17 out of the 22 compounds were very active at a concentration of 40 μM and, importantly, seven of them had an IC 50 on AA-triggered aggregation below 3 μM. The effects of the most active compounds were confirmed on collagen-triggered aggregation too. None of the tested compounds was toxic toward erythrocytes at 50 μM and four compounds partly inhibited proliferation of breast cancer cell line at 100 μM but not at 10 μM. Additionally, none of the compounds had a significant effect on blood coagulation or thrombin-triggered aggregation. This study hence reports four phenol derivatives (4-ethylcatechol, 4-fluorocatechol, 2-methoxy-4-ethylphenol and 3-methylcatechol) suitable for future in vivo testing.

Background: Antimicrobial resistance (AMR) has become an emerging threat worldwide, and developing countries such as Bangladesh are considered to be at greater risk of disseminating the resistant bacteria between human-animal interfaces., Objectives: The present study was carried out to determine the prevalence and AMR profile of Escherichia coli isolated from broiler chickens, the environment, and farmworkers. This study also aimed to identify the risk factors associated with multidrug-resistant (MDR) E. coli infection in broiler chickens. In addition, the presence of carbapenem resistance gene (NDM-1) was assessed., Methods: A total of 114 E. coli isolates, recovered from 150 samples (cloacal swabs = 50, farm sewage = 50, and hand washed water of farmworkers = 50) collected from 50 broiler farms, were identified by biochemical examination and polymerase chain reaction (PCR) assay. Antimicrobial susceptibility test was performed for 10 antibiotics by disk diffusion test. Carbapenem resistance gene (NDM-1) was detected by PCR. Risk factors were identified through multivariable logistic regression., Results: The highest prevalence of E. coli was recorded in broiler chickens (86%) and the lowest in farmworkers (66%). For MDR E. coli infection, 'winter season', 'absence of specific shoes for staff', and 'use of antibiotics without veterinarian's prescription' were the significant risk factors. High resistance of the E. coli isolates was observed against levofloxacin (81.6%), doxycycline (78.1%), cefotaxime (78.1%), and ciprofloxacin (70.2%). About 76% of the isolates demonstrated MDR. None of the isolates were positive for the NDM-1 gene., Conclusions: The high level and similar pattern of antibiotic resistance in E. coli isolates from broiler chickens, farmworkers, and sewage in poultry farms indicates a good possibility of spreading the antibiotic-resistant E. coli in such settings., (© 2021 The Authors. Veterinary Medicine and Science published by John Wiley & Sons Ltd.)

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., Competing Interests: No Conflict of Interest is declared, (Copyright (c) 2021 Sudipta Talukder, Md. Mehedi Hasan, Amit Kumar Mandal, Syeda Tanjina Tasmim, Mst. Sonia Parvin, Md. Yamin Ali, Azimun Nahar, Md. Zahorul Islam, Md. Taohidul Islam.)

Background and Aim: Subclinical mastitis (SCM) is an economically important disease for dairy cattle worldwide; therefore, regular screening is imperative to detect SCM at an early stage so as to control it. The study was conducted to compare the test characteristics of sodium lauryl sulfate (SLS) as a test reagent to detect SCM in dairy cows., Materials and Methods: First, 106 milk samples of dairy cows were subjected to available indirect screening tests (white side test [WST], surf field mastitis test, Leucocytest, and Immucell) considering somatic cell count (SCC) as gold standard test. Then 273 milk samples were allowed to react with different concentrations of SLS with or without sodium hydroxide (NaOH) and indicators (bromothymol blue [BTB] and bromocresol purple [BCP])., Results: SLS with or without NaOH yielded best reaction with the milk samples similar to Leucocytest. It was observed that the reaction of milk samples with SLS added with indicators (BTB and BCP) was easier to visualize than without indicators. SLS 3%+NaOH 1.5% with BTB and SLS 2% with BCP had high sensitivity, specificity, and substantial agreement with SCC. The area under the receiver operating characteristics curve of SLS 2% with BCP and SLS 3%+NaOH 1.5% with BTB was 0.917 and 0.875, respectively., Conclusion: It may be concluded that SLS 3%+NaOH 1.5% with BTB and SLS 2% with BCP may be the potential reagents for the development of an effective cow-side test to detect SCM, as the main ingredient SLS is considerably cheap and readily available in developing countries., (Copyright: © Tanni, et al.)

Infections by methicillin-resistant Staphylococcus aureus (MRSA) are continuously expanding within the community. Chicken meat is usually contaminated by MRSA, and this contaminated chicken meat is an important source of foodborne infections in humans. In this study, a cross-sectional supershop survey was conducted to determine the prevalence and antimicrobial resistance pattern of MRSA in 113 domestic frozen chicken meat samples purchased from nine branded supershops available in five divisional megacities of Bangladesh. The study also focused on the determination of methicillin resistance gene in MRSA isolates. S. aureus was identified by standard culture-based and molecular methods, and subjected to antimicrobial susceptibility testing. MRSA was screened by cefoxitin disk diffusion test. Methicillin resistance gene was identified by PCR. Of samples, 54.9% were positive for S. aureus , and, of these, 37.1% isolates were identified as MRSA. All the isolates were multidrug resistant (MDR): 52.2% were resistant to 6-8 antimicrobial classes, and 47.8% isolates to 9-12 classes. Three (3.2%) isolates of S. aureus were possible extensively drug resistant. The highest rates of resistance were observed against cefoxitin (100%), followed by nalidixic acid, ampicillin and oxacillin (97.7%), colistin (91.3%), amoxicillin-clavulanic acid and amoxicillin (87%), penicillin-G and cloxacillin (82.6%), oxytetracycline (78.3%), and cefixime (73.9%). Screening of methicillin resistance gene revealed that 43.5% isolates of MRSA were positive for mecA gene. The high prevalence of MDR MRSA in frozen chicken meat samples in this study emphasizes the need for better sanitary education of food handlers in hygienic practices focusing on their potential role as reservoirs and spreaders of MRSA.

Abstract: Salmonella is an important foodborne pathogen that causes public health problems globally, and the increase of antimicrobial resistance in Salmonella has intensified the problem. Chicken meat is an important reservoir and disseminator of Salmonella to humans. This study aimed at estimating the burden of Salmonella carrying extended-spectrum β-lactamase (ESBL) and their antimicrobial resistance pattern in 113 domestic frozen chicken meat samples purchased from supershops available in five divisional megacities of Bangladesh. The study also focused on the determination of β-lactamase-, and plasmid-mediated quinolone resistance-encoding genes. All samples were analyzed for the presence of Salmonella using selective media and PCR assay. Antimicrobial susceptibility test was done by disk diffusion test, and ESBL screening was performed by double-disk synergy tests. Resistance genes were detected using multiplex PCR. Of samples, 65.5% were positive for Salmonella spp., and, of these, 58.1% isolates were ESBL producers. All the isolates were multidrug resistant (MDR): 40.5% were resistant to both three to five and six to eight antimicrobial classes; 17.6% were resistant to 9 to 11 classes, and 1.4% isolates to 12 to 15 classes. The highest rates of resistance were observed against oxytetracycline (100%), followed by trimethoprim-sulfamethoxazole (89.2%), tetracycline (86.5%), nalidixic acid (83.8%), amoxicillin (74.3%), and pefloxacin (70.3%). Notably, 48.6% of isolates demonstrated resistance to imipenem. One (1.4%) isolate was possibly extensively drug resistant. All the isolates were positive for the blaTEM gene, 2.7% were positive for blaCTX-M-1, and 20.3% for blaNDM-1. The prevalence of qnrA and qnrS genes was 4.1 and 6.8%, respectively. This study shows that ESBL-producing Salmonella are widespread in frozen chicken meat in Bangladesh, which puts greater responsibility on food processors and policy makers to ensure food safety., (Copyright ©, International Association for Food Protection.)

Background and Aim: Subclinical mastitis (SCM) is one of the most prevalent diseases of dairy cows, and somatic cell count (SCC) is widely used to determine SCM and milk quality. However, setting the threshold of SCC is very important. This study aimed to determine the cow-level threshold of SCC to differentiate milk of SCM affected cows from normal milk in Bangladesh., Materials and Methods: Sensitivity (Se) and specificity (Sp) along with other characteristics of different thresholds of SCC were determined considering the bacterial culture as the gold standard test. Three definitions of intramammary infection (IMI) were set based on the group of pathogens involved. Five categories of SCC thresholds were considered for analysis., Results: Se and Sp of thresholds of SCC greatly varied as definitions of IMI changed. Irrespective of SCC thresholds, Se increased when isolation of major pathogens included in IMI definitions. Se decreased when SCC thresholds increased (from 100 to 300 × 10 3 cells/mL) for all IMI definitions and ranged from 60.6% to 20.3%. The highest Se was found at low SCC threshold (100 × 10 3 cells/mL), which resulted in less false-negative outcome. On the other hand, Sp increased with the increment of SCC thresholds giving rise to a less false-positive outcome., Conclusion: The cow-level SCC threshold of 100 × 10 3 cells/mL was found appropriate to identify subclinical IMI of dairy cows., (Copyright: © Sumon, et al.)

Campylobacter spp. is one of the most frequent causes of foodborne gastroenteritis. This study aimed to estimate the prevalence and to identify the risk factors of farm-level Campylobacter infection in meat-type chicken flocks. A cross-sectional study was conducted in two selected districts of Bangladesh over the period of January to July 2019. A total of 84 pooled cloacal swab samples were collected from 84 broiler and cockerel farms. Data on farm management, biosecurity, and hygiene practices were collected using a structured questionnaire through a face-to-face interview during sampling. Thereafter, Campylobacter spp. were isolated through bacteriological culture and identified by Gram staining and biochemical tests. Furthermore, the isolates were confirmed using the polymerase chain reaction by targeting the 16S rRNA gene. The risk factors were analyzed at the farm level using multivariable logistic regression with the significant levels of P-value ≤ 0.05. Among the 84 farms, 34 were positive to Campylobacter spp.; thus, the prevalence was estimated to be 40.5% (95% CI: 30.1%-51.8%). In risk factor analysis, the following factors were found to be significantly associated with Campylobacter infection: shed older than five years, birds older than 30 days, flock size with more than 1500 birds, downtime less than seven days, no disinfection of shed surroundings during rearing, rice husk as litter materials, and less than 10 years of farming experience. The study identified the factors that could lead to the setting of effective interventions in controlling Campylobacter infection in chickens to reduce campylobacteriosis in humans through meat consumption., Competing Interests: Declaration of Competing Interest None., (Copyright © 2020 Elsevier B.V. All rights reserved.)

E scherichia coli is known as one of the most important foodborne pathogens in humans, and contaminated chicken meat is an important source of foodborne infection with this bacterium. The occurrence of extended-spectrum β-lactamase (ESBL)-producing E. coli (ESBL-Ec), in particular, in chicken meat is considered a global health problem. This study aimed to determine the magnitude of E. coli , with special emphasis on ESBL-Ec, along with their phenotypic antimicrobial resistance pattern in frozen chicken meat. The study also focused on the determination of ESBL-encoding genes in E. coli . A total of 113 frozen chicken meat samples were purchased from 40 outlets of nine branded supershops in five megacities in Bangladesh. Isolation and identification of E. coli were done based on cultural and biochemical properties, as well as PCR assay. The resistance pattern was determined by the disc diffusion method. ESBL-encoding genes were determined by multiplex PCR. The results showed that 76.1% of samples were positive for E. coli , of which 86% were ESBL producers. All the isolates were multidrug-resistant (MDR). Resistance to 9-11 and 12-13 antimicrobial classes was observed in 38.4% and 17.4% isolates, respectively, while only 11.6% were resistant to 3-5 classes. Possible extensive drug resistance (pXDR) was found in 2.3% of isolates. High single resistance was observed for oxytetracycline (93%) and amoxicillin (91.9%), followed by ampicillin (89.5%), trimethoprim-sulfamethoxazole, and pefloxacin (88.4%), and tetracycline (84.9%). Most importantly, 89.6% of isolates were resistant to carbapenems. All the isolates were positive for the bla TEM gene. However, the bla SHV and bla CTX-M-2 genes were identified in two ESBL-non producer isolates. None of the isolates carried the bla CTX-M-1 gene. This study provided evidence of the existence of MDR and pXDR ESBL-Ec in frozen chicken meat in Bangladesh, which may pose a risk to human health if the meat is not properly cooked or pickled raw only. This emphasizes the importance of the implementation of good slaughtering and processing practices by the processors., Competing Interests: The authors declare that they have no competing interests. The funders had no role in the study design, data collection and analysis, manuscript preparation, or decision to publish the manuscript.

Objective: The influence of intramammary infection (IMI) and types of bacteria was assessed on somatic cell count (SCC) in dairy cows' milk with respect to breed, age, parity, stage of lactation, milk production, and mammary quarter location., Materials and Methods: After recording data in a structured questionnaire, 360 samples of quarter milk were collected. The samples were subjected to SCC and isolation and identification of bacteria. The data were analyzed to find out the significant influence of independent factors on SCC and IMI., Results: The infected quarters had a significantly higher mean SCC (210.52 × 10 3 cells/ml) compared to uninfected ones (32.72 × 10 3 cells/ml). The mean SCC was the highest for IMI with Enterobacter spp. (338.00 × 10 3 cells/ml) followed by Bacillus spp. (319.20 × 10 3 cells/ml), coagulase-negative Staphylococci (CNS) (268.17 × 10 3 cells/ml), Staphylococcus aureus (218.31 × 10 3 cells/ml), and Escherichia coli (200.75 × 10 3 cells/ml) and the lowest for Pseudomonas aeruginosa (66.33 × 10 3 cells/ml). Milk of rear quarters had a significantly higher SCC than the front quarters. SCC increased with increasing age, parity, and lactation stage regardless of whether cows are infected or not. The IMI was more prevalent in rear quarters (42.2%) and cows at early (≤7 days) lactation (100.0%). Cows having a parity of ≥5 and crossbred and high yielding (>5 l) cows had also a higher rate of IMI of 38.2%, 36.7%, and 38.2%, respectively., Conclusion: The IMI and type of bacteria were the principal factors for SCC variation. Besides, mammary quarter location, age, and parity should be taken into consideration during the interpretation of SCC., Competing Interests: The authors declare that they have no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (Copyright: © Journal of Advanced Veterinary and Animal Research.)

Objective: The study was aimed to analyze the microbiological quality of mixed vegetable salads and to understand the risk related with its consumption from different restaurants around Bangladesh Agricultural University campus in Mymensingh., Materials and Methods: Sixty (60) samples of mixed vegetable salads were taken from twelve (12) different restaurants in five different time points from each restaurant. In parallel, restaurant workers were asked for handling practices while the consumers were interviewed about their salad consumption pattern and whether they had experienced any health-related problems. Microbial risk assessment of Staphylococcus spp., Salmonella spp., and Escherichia coli was estimated by Monte Carlo simulation (10,000 iterations), an exponential model., Results: Aerobic plate count was ranged from 7.73 ± 0.61 to 9.04 ± 0.26 log cfu/gm, Staphylococcus spp. from 4.64 ± 0.61 to 6.42 ± 0.53 log cfu/gm, Salmonella spp. from 4.75 ± 0.08 to 5.27 ± 0.53 log cfu/gm, and E. coli from 4.98 ± 0.20 to 6.66 ± 0.80 log cfu/gm. From the survey, it was found that total consumers had 18% chances where the male had 13% and the female had 30% chances of being infected with salads. Again frequent, average, and occasional consumers had 31%, 13%, and 0% chances, respectively, of being infected with those salads. From the Monte Carlo simulation, the calculated mean annual risks of Staphylococcus spp., Salmonella spp., and E. coli infection for the three exposure scenarios were found to be about 100%., Conclusion: The study actually revealed the potential microbial contamination in mixed vegetable salads which may impact on food safety and human health. So, the findings suggest that following hygienic measures during processing and handling the microbiological quality of mixed vegetables salads can be improved., Competing Interests: The authors declare that there is no conflict of interest toward the publication of this article., (Copyright: © Journal of Advanced Veterinary and Animal Research.)

Background: The various parts of Cresecentia cujete have some important biological activities. In folklore medicine leaves are used to treat hematomas, tumors and hypertension. Fruit decoction is used to treat diarrhea, stomachaches, cold, bronchitis, cough, asthma, and urethritis. The present study was designed to explore the anti-inflammatory and antibacterial potential of C. cujete leaves and stem bark. Anti-inflammatory activity was evaluated by in vitro human red blood cell (HRBC) membrane stabilization method and antibacterial activity by disc diffusion method., Methods: In vitro anti-inflammatory activity was evaluated by human red blood cell (HRBC) membrane stabilization method while in vitro antibacterial activity was evaluated using cultures of Escherichia coli and Staphylococcus aureus by disc diffusion method. Total phenolic (TPC) and total flavonoid contents (TFC) of the crude extract and fractions were also determined by Folin-Ciocalteu's phenol reagent and by aluminium chloride method, respectively., Results: The crude ethanol extract (CEE) of leaves and bark (concentration of each 1.0 mg/ml) demonstrated strong membrane stabilizing activity (53.86 and 61.85% protection, respectively), whereas their chloroform fractions (CHF) revealed moderate activity (48.74 ± 0.56 and 43.55 ± 6.20 %, respectively) compared with standard aspirin (concentration 0.10 mg/ml) which showed 75.81% protection in this test. All the samples showed a dose dependent anti-inflammatory activity in HRBC membrane stabilization test. Total phenolic (TPC) and total flavonoid contents (TFC) of the crude extract and fractions were also determined. Again, in in vitro antibacterial study, the extractives exhibited potent antibacterial activity., Conclusion: Results from this study showed that the leaves and bark of C. cujete possessed anti-inflammatory as well as antibacterial activities indicating that the plant extract has therapeutic potential against the bacterial infection and also have effect on disease processes by causing destabilization of biological membranes.

Background: Tribolium castaneum (Herbst) is a harmful pest of stored grain and flour-based products in tropical and subtropical region. In the present study, rhizome of Drynaria quercifolia (J. Smith) was evaluated for pesticidal and pest repellency activities against T. castaneum, using surface film method and filter paper disc method, respectively. In addition, activity of the isolated compound 3,4-dihydroxybenzoic acid was evaluated against the pest., Results: Chloroform soluble fraction of ethanol extract of rhizome of D. quercifolia showed significant pesticidal activity at doses 0.88 to 1.77 mg/cm(2) and significant pest repellency activity at doses 0.94 to 0.23 mg/cm(2). No pesticidal and pest repellency activity was found for petroleum ether, ethyl acetate and methanol soluble fractions of ethanol extract as well as for 3,4-dihydroxybenzoic acid., Conclusion: Considering our findings it can be concluded that chloroform soluble fraction of rhizome of D. quercifolia is useful in controlling T. castaneum of stored grain and flour-based products.

Objective: To ascertain analgesic, antibacterial and central nervous system (CNS) depressant activities of ethyl acetate, dichloromethane and carbon tetrachloride fractions of methanol extract of Albizia procera (A. procera) leaves., Methods: Leaves extracts of A. procera were tested for analgesic activity by acetic acid induced and formalin test method in mice. The in vitro antibacterial activity was performed by agar well diffusion method. CNS depressant activity was evaluated by hole cross and open field tests., Results: All the extracts at 200 mg/kg exhibited significant (P<0.01) analgesic activity in acetic acid induced and formalin tests method in mice. Analgesic activity of the ethyl acetate fraction was almost same like as standard drug indomethacin in acetic acid induced method. The CNS depressant activity of the extracts at 500 mg/kg was comparable to the positive control diazepam as determined by hole cross and open field test method. The extracts exhibited moderate antimicrobial activity against all the tested microorganisms (Staphylococcus aureus, Bacillus cereus, Pseudomonas aeruginosa, Esherichia coli, Shigella soneii, Shigella boydii) at concentration of 0.8 mg/disc. The measured diameter of zone of inhibition for the extracts was within the range of 7 to 12 mm which was less than the standard kanamycin (16-24 mm)., Conclusions: It is concluded that all the extracts possess potential analgesic and CNS depressants activity. This study also showed that different fractions of methanol extract could be potential sources of new antimicrobial agents.

Background: Antioxidant compounds like phenols and flavonoids scavenge free radicals and thus inhibit the oxidative mechanisms that lead to control degenerative and other diseases. The aim of this study was to investigate the antioxidant activity in vitro, total phenolic and flavonoid contents in ethanol extracts and fractions of Crescentia cujete leaves and stem bark., Methods: Crescentia cujete leaves and bark crude ethanol extract (CEE) and their partitionates petroleum ether (PEF), chloroform (CHF), ethyl acetate (EAF) and aqueous (AQF) were firstly prepared. Different established testing methods, such as 1, 1-diphenyl-2-picryl hydrazyl (DPPH) radical, ferric reducing power (FRP), and total antioxidant capacity (TAC) assays were used to detect the antioxidant activity. Further, the total yield, total phenolic (TPC) and total flavonoid contents (TFC) of CEE and all the fractions were determined. Ethanol extracts of both leaves and stem bark were also subjected to preliminary phytochemical screening to detect the presence of secondary metabolites, using standard phytochemical methods (Thin layer chromatography and spray reagents)., Results: Phytochemical screening of crude ethanol extract of both leaves and stem bark revealed the presence of steroids, flavonoids, saponins, tannins, glycosides and terpenoids. All the fractions and CEE of leaves and bark exhibited antioxidant activities, however, EAF of leaves showing the highest antioxidant activity based on the results of DPPH, FRP and TAC assay tests. The above fraction has shown the significant DPPH scavenging activity (IC50 = 8.78 μg/ml) when compared with standard ascorbic acid (IC50 =7.68 μg/ml). The TAC and FRP activities increased with increasing crude extract/fractions content. The TPC (371.23 ± 15.77 mg GAE/g extract) and TFC (144.64 ± 5.82 mg QE/g extract) of EAF of leaves were found significantly higher as compared to other solvent fractions for both leaves and bark. TPC were highly correlated with the antioxidant activity (R2 = 0.9268 and 0.8515 in DPPH test for leaves and bark, respectively)., Conclusion: The results of the study show that leaves of C. cujete possesses significant free radical scavenging properties compared with stem bark and a clear correlation exists between the antioxidant activity and phenolic content.

Objective: To evaluate the pharmacological activity against diarrhea of methanol and petroleum ether extract of Desmodium puchellum (Family: Fabaceae) leaves., Methods: The extract was evaluated for castor oil-induced diarrhea and enteropooling as well as intestinal motility in rats. Both of the extracts were given to the rats at 200 mg/kg orally. Loperamide was used as a standard drug for diarrhea., Results: The diarrheal severity was reduced significantly (P<0.05) by methanol and petroleum ether extracts by 31.95% and 28.33%, respectively, whereas 54.75% inhibition was found for standard drug loperamide at 5 mg/kg. The two extracts also significantly (P<0.05) reduced the intestinal volume in case of castor oil induced enteropooling., Conclusions: It is concluded that both fractions contain some biologically active ingredients that are active for anti-diarrheal actions whereas methanol fraction has better potential.

The purpose of the study was to identify the potential risk factors for subclinical mastitis (SCM) in lactating dairy cows in Bangladesh. A cross-sectional study was carried out on randomly selected 212 smallholder dairy farms of Sadar upazilas of Rangpur, Mymensingh, and Satkhira districts of Bangladesh during January to October 2011. The direct interview using a structured questionnaire and physical examination of the cows were done to collect data on 15 variables. Milk samples collected from study cows were subjected to California Mastitis Test (CMT). The diagnosis of SCM was based on the results of CMT and physical examination of udder and milk. The bivariable followed by multivariable analysis was done using SPSS 17.0. Of the total cows examined, 20.2 % had subclinical mastitis. In bivariable analysis, eight risk factors were identified. However, in the final model of multivariable analysis, four potential risk factors were identified. These were history of previous clinical mastitis (odds ratio (OR) 10.51, p<0.001), pendulous type of udder (OR 2.26, p=0.008), no grass feeding (OR 1.84, p=0.039), and body condition score (BCS) 2.5 or less (OR 7.25, p=0.054). Four different factors were significantly associated with the occurrence of subclinical mastitis, which need to be considered in the control of the disease. However, particular emphasis should be given on grass feeding and BCS because these traits can be modified or improved to allow prevention of SCM.

Herbal medicines have traditionally been used worldwide for the prevention and treatment of liver disease with fewer adverse effects. The leaves of the Syzygium jambos (SJL) plant were chosen and studied for their antioxidant activity in vitro and hepatoprotective activity in vivo. The antioxidant activity of the ethanol extract was examined in vitro using a 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay, reducing capacity, total phenol, total flavonoid content, and total antioxidant capacity. The extract had significant dose-dependent antioxidant activity in all in vitro experiments. IC(50) values of SJL and ascorbic acid (standard) were found to be 14.10 and 4.87 μg/mL, respectively, according to a DPPH radical scavenging assay. Hepatoprotective activity of the plant extract was evaluated in a rat model of carbon tetrachloride (CCl(4))-induced liver damage. CCl(4) significantly altered serum marker enzymes, total bilirubin, total protein, and liver weight. The extract caused these values to return to normal in rats with CCl(4)-induced liver damage that were given SJL. This indicated the hepatoprotective potential of SJL and was comparable to use of the standard drug silymarin. Thus, the present study revealed that SJL may have antioxidant and hepatoprotective activity.

Zebrafish hoxb1b is expressed during epiboly in the posterior neural plate, with its anterior boundary at the prospective r4 region providing a positional cue for hindbrain formation. A similar function and expression is known for Hoxa1 in mice, suggesting a shared regulatory mechanism for hindbrain patterning in vertebrate embryos. To understand the evolution of the regulatory mechanisms of key genes in patterning of the central nervous system, we examined how hoxb1b transcription is regulated in zebrafish embryos and compared the regulatory mechanisms between mammals and teleosts that have undergone an additional genome duplication. By promoter analysis, we found that the expression of the reporter gene recapitulated hoxb1b expression when driven in transgenic embryos by a combination of the upstream 8.0-kb DNA and downstream 4.6-kb DNA. Furthermore, reporter expression expanded anteriorly when transgenic embryos were exposed to retinoic acid (RA) or LiCl, or injected with fgf3/8 mRNA, implicating the flanking DNA examined here in the responsiveness of hoxb1b to posteriorizing signals. We further identified at least two functional RA responsive elements in the downstream DNA that were shown to be major regulators of early hoxb1b expression during gastrulation, while the upstream DNA, which harbors repetitive sequences with apparent similarity to the autoregulatory sequence of mouse Hoxb1, contributed only to later hoxb1b expression, during somitogenesis. Possible implications in vertebrate evolution are discussed based on these findings., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Zebrafish pou5f1, also known as pou2, encodes a POU-family transcription factor that is transiently expressed in the prospective midbrain and anterior hindbrain during gastrulation, governing brain development. In the present study, we found that the main regulatory elements reside in the proximal upstream DNA sequence from -2.2 to -0.12 kb (the -2.2/-0.1 region). The electrophoretic gel mobility shift assay (EMSA) revealed four functional octamer sequences that can associate with zebrafish Pou2/Pou5f1. The expression of mutated reporter constructs, as well as EMSA, suggested that these four octamer sequences operate in a cooperative manner to drive expression in the mid/hindbrain. We also identified a retinoic acid (RA)-responsive element in this proximal region, which was required to repress transcription in the posterior part of the embryo. These data provide a scheme wherein pou2/pou5f1 expression in zebrafish embryos is regulated by both an autoregulatory loop and repression by RA emanating from the posterior mesoderm.

Fgf8 is expressed in the isthmic region of the developing brain, serving an organizing function in vertebrate embryos. We previously identified S4.2 downstream to the zebrafish fgf8 gene as a regulatory region that drives transcription in the anterior hindbrain. Here, we investigated the mechanism of fgf8 regulation by the S4.2 region during development. Reporter analyses in embryos revealed that S4.2 closely recapitulates fgf8 expression in the anteriormost hindbrain during somitogenesis. This region contains a sequence highly conserved in fgf8 of diverse vertebrates. Further analyses of S4.2 revealed a 342-bp core region composed of three subregions (#2, #3, and #4). Regions #3 and #4 drove expression broadly in the brain from the midbrain to r5 of the hindbrain, whereas a 28-bp sequence in #2 repressed ectopic expression in the midbrain and in r2 to r5. The enhancer function of S4.2 was absent in pax2a mutant embryos, while it was activated ectopically by pax2a misexpression in the hindbrain. We identified two sites in the core region that are bound by Pax2a in vitro and in vivo, the disruption of which abrogated the S4.2 activity. Thus, fgf8 expression in the anteriormost hindbrain involves activation and repression, with Pax2a as a pivotal regulator.