BCAAs acutely drive glucose dysregulation and insulin resistance: role of AgRP neurons.

BACKGROUND: High-protein diets are often enriched with branched-chain amino acids (BCAAs) known to enhance protein synthesis and provide numerous physiological benefits, but recent studies reveal their association with obesity and diabetes. In support of this, protein or BCAA supplementation is shown to disrupt glucose metabolism while restriction improves it. However, it is not clear if these are primary, direct effects of BCAAs or secondary to other physiological changes during chronic manipulation of dietary BCAAs. METHODS: Three-month-old C57Bl/6 mice were acutely treated with either vehicle/BCAAs or BT2, a BCAA-lowering compound, and detailed in vivo metabolic phenotyping, including frequent sampling and pancreatic clamps, were conducted. RESULTS: Using a catheter-guided frequent sampling method in mice, here we show that a single infusion of BCAAs was sufficient to acutely elevate blood glucose and plasma insulin. While pre-treatment with BCAAs did not affect glucose tolerance, a constant infusion of BCAAs during hyperinsulinemic-euglycemic clamps impaired whole-body insulin sensitivity. Similarly, a single injection of BT2 was sufficient to prevent BCAA rise during fasting and markedly improve glucose tolerance in high-fat-fed mice, suggesting that abnormal glycemic control in obesity may be causally linked to high circulating BCAAs. We further show that chemogenetic over-activation of AgRP neurons in the hypothalamus, as present in obesity, significantly impairs glucose tolerance that is completely normalized by acute BCAA reduction. Interestingly, most of these effects were demonstrated only in male, but not in female mice. CONCLUSION: These findings suggest that BCAAs per se can acutely impair glucose homeostasis and insulin sensitivity, thus offering an explanation for how they may disrupt glucose metabolism in the long-term as observed in obesity and diabetes. Our findings also reveal that AgRP neuronal regulation of blood glucose is mediated through BCAAs, further elucidating a novel mechanism by which brain controls glucose homeostasis.

Revisiting the Role of Vitamins and Minerals in Alzheimer's Disease.

Alzheimer's disease (AD) is the most common type of dementia that affects millions of individuals worldwide. It is an irreversible neurodegenerative disorder that is characterized by memory loss, impaired learning and thinking, and difficulty in performing regular daily activities. Despite nearly two decades of collective efforts to develop novel medications that can prevent or halt the disease progression, we remain faced with only a few options with limited effectiveness. There has been a recent growth of interest in the role of nutrition in brain health as we begin to gain a better understanding of what and how nutrients affect hormonal and neural actions that not only can lead to typical cardiovascular or metabolic diseases but also an array of neurological and psychiatric disorders. Vitamins and minerals, also known as micronutrients, are elements that are indispensable for functions including nutrient metabolism, immune surveillance, cell development, neurotransmission, and antioxidant and anti-inflammatory properties. In this review, we provide an overview on some of the most common vitamins and minerals and discuss what current studies have revealed on the link between these essential micronutrients and cognitive performance or AD.

A Cross-Sectional Seroepidemiological Study on Infectious Bursal Disease in Backyard Chickens in the Mymensingh District of Bangladesh.

Infectious bursal disease (IBD) is a highly contagious disease that causes significant economic loss in chickens. A cross-sectional study was conducted in the Mymensingh district of Bangladesh to determine the seroprevalence of IBD virus (IBDV) antibodies in backyard chickens and their association with different epidemiological risk factors. A total of 460 serum samples were randomly collected from backyard chickens that had not been previously vaccinated against IBDV. The collected sera were examined using an enzyme-linked immunosorbent assay (ELISA). Data on epidemiological risk factors were collected through face-to-face interviews with owners and subjected to both uni- and multivariable risk analyses to determine their association with IBDV infection. Using ELISA, the overall seroprevalence of IBDV antibodies in backyard chickens was 83.4% (95% confidence interval: 79.8%-86.6%), among which, a significantly higher seroprevalence was recorded in females (83.4%, 345/350), 4-6 weeks age group (95.3%, 244/256), and unhealthy (95.0%, 57/60) backyard chickens than those of males, other age groups, and healthy chickens, respectively. Furthermore, chickens reared in free-ranging housing systems (93.3%, 280/300) and poor-conditioned houses (98.0%, 147/150) showed a significantly higher seropositivity of IBDV antibodies than those reared in separated housing systems and other hygienic-conditioned houses, respectively. Moreover, compared with their counterparts, a higher but nonsignificant seroprevalence of IBDV antibodies was observed in backyard chickens that were selected from Fulbaria Upazila (88.8%; 80/90) and which were brought from the marketplace (85.7%, 60/70). A higher seropositivity of IBDV antibodies was shown to be statistically associated with various critical epidemiological risk factors, indicating that field strains of IBDV were exposed in backyard chickens and could be readily transferred horizontally. Proper prevention and control methods, villagers' awareness of IBD, and the rapid and widespread use of seroepidemiological investigations could help to reduce the spread of IBDV infection in backyard chickens.

Higher seasonal temperature enhances the occurrence of methicillin resistance of Staphylococcus aureus in house flies (Musca domestica) under hospital and environmental settings.

Antimicrobial resistance (AMR) emergence in commensal and pathogenic bacteria is a global health issue. House flies (Musca domestica) are considered as biological and mechanical vectors for pathogens causing nosocomial infections, including methicillin-resistant Staphylococcus aureus (MRSA). However, the prevalence of antimicrobial resistance and the role of temperature on the occurrence of Staphylococcus aureus and MRSA in house flies in a hospital environment have not been studied. A total of 400 house flies were collected in winter and summer from four hospital-associated areas in Mymensingh, Bangladesh. Detection of S. aureus and MRSA in flies was done by culturing, staining, and PCR methods targeting nuc and mec genes (mecA and mecC), respectively. Disc diffusion test was used to detect resistance phenotype against six antimicrobials. Logistic regression models were constructed to assess the effect of temperature on the frequency of antimicrobial resistance, and on the presence of the nuc and mecA genes, and location of samples in and around a hospital environment. By PCR, S. aureus was detected in 208 (52%) samples. High frequencies of resistance (≥ 80% of isolates) to amoxicillin, azithromycin, and oxacillin were observed by disk diffusion test. Increase in temperature had a positive effect on the occurrence of S. aureus and MRSA isolates as well as on their resistance to individual and multiple antimicrobials. Among the study areas, hospital premises had increased odds of having S. aureus. Increased temperature of summer significantly increased the occurrence of MRSA in house flies in and around the hospital environment, which might pose a human and animal health risk.

groEL gene-based molecular detection and antibiogram profile of Riemerella anatipestifer from duck in Bangladesh.

Objectives: This study was designed to detect Riemerella anatipestifer through polymerase chain reaction (PCR) from duck farming areas of the Mymensingh and Sylhet divisions and to determine the antibiogram profile of the PCR-positive isolates using the disc diffusion method. Materials and Methods: Fifty two samples were collected, comprising clinically sick (32 ducks) and dead ducks (20). PCR confirmation was accomplished, and consistent findings were observed, employing R. anatipestifer groEL (271-bp) gene as appropriate molecular markers. For further clarification, see R. anatipestifer specific PCR assay (546-bp) and gyrB-based PCR (162-bp) were also done. The disc diffusion method was followed for the antibiotic susceptibility test of the isolates against commonly used antibiotics. Results: A total of 21 samples, 8 from clinically sick birds and 13 from dead birds, showed positive results in both conventional and molecular assays out of 52 samples. High occurrences were found in oropharyngeal swabs from sick ducks and the liver and heart from dead ducks. Antibiotic susceptibility testing revealed that the isolates were 100% resistant to penicillin G, cefradine, streptomycin, neomycin, gentamycin, meropenem, and erythromycin, but 100% sensitive to -cotrimoxazole, florfenicol, and levofloxacin. Conclusion: For diverse duck-populated areas in Bangladesh, this study shows the severity of R. anatipestifer infection among ducks.

Seroprevalence and risk factors of avian reovirus in backyard chickens in different areas of Mymensingh district in Bangladesh.

OBJECTIVE: The present study estimated the seroprevalence of avian reovirus (ARV) infections in backyard chickens of the Mymensingh district in Bangladesh. MATERIALS AND METHODS: Considering several risk factors, a total of 460 serum samples were collected from backyard chickens from eight Upazilas of the Mymensingh district in Bangladesh. Blood samples were taken from the wing vein using 3-ml sterile syringes and kept at room temperature for clotting in a slanting position and then transported to the laboratory maintaining the cool chain. Subsequently, the prepared sera were harvested and stored at -20°C until used. Finally, an indirect enzyme-linked immunosorbent assay (ELISA) was performed to detect ARVspecific antibodies using a commercial ARV antibody detection ELISA test kit. RESULTS: The results revealed high prevalence rates of ARV antibodies, with a total seroprevalence of 69.78% (321/460). Area-wise, 74.55% (82/110) seroprevalence was recorded as the highest in Mymensingh Sadar, whereas 64% (32/50) was the lowest in Gauripur Upazila. With regard to sex, female chickens showed a significantly higher (p < 0.05) seroprevalence as 90.33% (271/300) compared to male chickens 31.25% (50/160). With regard to age groups, the seroprevalence of ARV infection was 59.33% (89/150) within 2-8 weeks, 82% (205/250) within 9-16 weeks, and 45% (27/60) within 17-20 weeks, respectively. Based on hygienic conditions, the highest seroprevalence of ARV was noted in backyard chickens housed in poor conditions 80% (120/150) than good conditions 50% (40/80). Backyard chickens reared in free-ranging conditions exhibited a significantly higher seroprevalence 73.33% (220/300) of ARV antibodies compared to rearing in separate houses 63.12% (101/160). The seroprevalence of ARV was higher in crossbreeds 71.67% (43/60), brought from market 76% (38/50), and unhealthy 78.57% (55/70) backyard chickens than non-descriptive indigenous 69.5% (278/400), home-reared 69.02% (283/410), and healthy chickens 68.21% (266/390). CONCLUSION: The high prevalence of ARV antibodies revealed in the current study indicates an extensive exposure of ARV to backyard chickens in Bangladesh that may be transmitted naturally to other chickens, ultimately leading to ominous economic effects on the poultry sector.

Detection of antibiotic-resistant bacteria and their resistance genes from houseflies.

BACKGROUND AND AIM: Houseflies (Musca domestica) are synanthropic insects which serve as biological or mechanical vectors for spreading multidrug-resistant bacteria responsible for many infectious diseases. This study aimed to detect antibiotic-resistant bacteria from houseflies, and to examine their resistance genes. MATERIALS AND METHODS: A total of 140 houseflies were captured using sterile nylon net from seven places of Mymensingh city, Bangladesh. Immediately after collection, flies were transferred to a sterile zipper bag and brought to microbiology laboratory within 1 h. Three bacterial species were isolated from houseflies, based on cultural and molecular tests. After that, the isolates were subjected to antimicrobial susceptibility testing against commonly used antibiotics, by the disk diffusion method. Finally, the detection of antibiotic resistance genes tetA, tetB, mcr-3, mecA, and mecC was performed by a polymerase chain reaction. RESULTS: The most common isolates were Staphylococcus aureus (78.6%), Salmonella spp., (66.4%), and Escherichia coli (51.4%). These species of bacteria were recovered from 78.3% of isolates from the Mymensingh Medical College Hospital areas. Most of the isolates of the three bacterial species were resistant to erythromycin, tetracycline, penicillin and amoxicillin and were sensitive to ciprofloxacin, ceftriaxone, chloramphenicol, gentamicin, and azithromycin. Five antibiotic resistance genes of three bacteria were detected: tetA, tetB, mcr-3, and mecA were found in 37%, 20%, 20%, and 14% isolates, respectively, and no isolates were positive for mecC gene. CONCLUSION: S. aureus, Salmonella spp., and E. coli with genetically-mediated multiple antibiotic resistance are carried in houseflies in the Mymensingh region. Flies may, therefore, represent an important means of transmission of these antibiotic-resistant bacteria, with consequent risks to human and animal health.

Microbial risk assessment of ready-to-eat mixed vegetable salads from different restaurants of Bangladesh Agricultural University campus.

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.

Microbial load in bio-slurry from different biogas plants in Bangladesh.

OBJECTIVE: The study was aimed to isolate, identify, and characterize common indicator bacteria, including Escherichia coli, Salmonella spp., and Staphylococcus spp. in manure and bio-slurry samples of different livestock farms and biogas plants of Bangladesh. MATERIALS AND METHODS: A total of 114 samples of manure and bio-slurry were collected from different livestock farms and biogas plants in Bangladesh. The total viable count (TVC), E. coli, Salmonella spp., and Staphylococcus spp. counts were determined by the spread plate technique method. Isolation and identification were performed by colony characteristics, staining, biochemical tests, and, finally, by using PCR. Antibiotic susceptibility test of the isolated bacteria was tested against commonly used antibiotics by using the disk diffusion method. RESULTS: The mean TVC, E. coli, Salmonella spp., and Staphylococcus spp. counts were ranged from 8.19-10.75, 5.2-6.96, 5.81-6.87, 5.68-7.68 in manure samples and 7.26-8.65, 3.82-5.2, 4-5.54, 3.14-5.9 log cfu/gm in bio-slurry, respectively. In anaerobic digester after 30 days digestion, the presence of E. coli, Salmonella spp., and Staphylococcus spp. varied from 0-5.11, 0-4.84, and 0-5.59 log cfu/gm at 25°C, 27°C, 29°C, and 45°C temperature. Above-mentioned bacteria were absent in bio-slurry collected from anaerobic digester after 60 days digestion at environmental temperature. Bacterial counts were reduced significantly in both household slurry pits and experimental anaerobic digester. Antibiotic susceptibility results revealed that multidrug-resistant indicator bacteria were present in the bio-slurry samples. CONCLUSION: Our findings conclude that the microbial load after treatment of animal manure via anaerobic digestion (Biogas plant) was grossly reduced and the reduction of bacterial pathogen depends on the duration and temperature of digestion.

Serological detection of avian reovirus in different poultry flocks of Gazipur and Mymensingh districts of Bangladesh.

BACKGROUND AND AIM: Avian reovirus (ARV) is a constraint to poultry industry in Bangladesh as a cause of several diseases in chickens, especially in broiler. However, the actual status of the viral infection is not known because the large-scale study is not conducted in this country. Therefore, this study aimed to check the presence and distribution of ARV-specific antibody in respect to area, types of chickens (broiler breeder, broiler, and layer), vaccination status, and age of chickens in Gazipur and Mymensingh districts of Bangladesh. MATERIALS AND METHODS: A total of 276 chickens' blood samples were collected from two well-organized broiler breeder stock, seven broiler farms, and five layer farms located at two districts, namely Gazipur and Mymensingh of Bangladesh. Blood samples were collected from wing vein of the apparently healthy chickens using 3 ml of syringe and serum was harvested by keeping the syringe at room temperature in slanting position. The sera were transferred to the laboratory by maintaining the cool chain and further processing was performed by indirect enzyme-linked immunosorbent assay using ARV antibody test kit. RESULTS: The results of serological test revealed that an average of 39.5% seropositive against ARV was recorded in chickens of Gazipur and Mymensingh districts. Among these, chickens of Gazipur district had the highest seropositivity of 50.5% than Mymensingh (30.7%). With respect to vaccination status, the seropositivity of vaccinated chickens in both areas was 100% and non-vaccinated chickens was 50.5% in Gazipur and 30.7% in Mymensingh district, respectively. However, regarding age groups, the seropositivity was higher in the age of 4-6 weeks (64.5%). CONCLUSION: The present serological findings showed a higher prevalence of ARV-specific antibodies in broiler birds. It indicates that the poultry industries of Bangladesh are contaminated with ARV which may naturally be transmitted to chickens either vertically or horizontally.

Emergence of colistin-resistant Escherichia coli in poultry, house flies, and pond water in Mymensingh, Bangladesh.

OBJECTIVE: Emergence of colistin-resistant Escherichia coli (CREC) has generated a sense of public alarm. The objective of this study was to detect the CREC and identification of the gene responsible for such resistance. MATERIALS AND METHODS: A total of 150 samples comprising poultry cloacal swab, house flies (Musca domestica), and pond water were collected randomly from Mymensingh, Bangladesh and analyzed. Isolation and identification of E. coli were done based on culture and E. coli 16S rRNA gene-specific polymerase chain reaction (PCR). Phenotypic detection of CREC was done by disk diffusion method. Finally, colistin resistance genes were detected by PCR by using colistin resistant gene mcr3 specific primers. RESULTS: Among the 150 samples, phenotypically 18.00% (n = 27/150) isolates were found as colistin resistant. By PCR, 8.00% of the E. coli isolates were found positive for the presence of mcr3 gene. CONCLUSIONS: Colistin resistant E. coli carrying mcr3 are detected in poultry, house flies and water that are of great public health concern.

Characterization of Staphylococcus aureus isolated from chicken and quail eggshell.

OBJECTIVES: This study was conducted to assess the prevalence and characterization of Staphylococcus aureus from chicken and quail eggshells and to study the antibiogram of the isolates. MATERIALS AND METHODS: A total of 300 eggs (220 chicken eggs and 80 quail eggs) were collected from different retail shops and farms in Mymensingh district. Swabs taken from the egg surfaces were cultured on Mannitol Salt Agar for the isolation of S. aureus. Polymerase chain reaction was conducted for confirmatory identification of the bacterial species targeting nuc gene, followed by confirmation of methicillin-resistant S. aureus by targeting the mecA gene. Antibiotic sensitivity test of the isolated bacteria was done against commonly used antibiotics by the disk diffusion method. RESULTS: The prevalence of Staphylococcus spp. and S. aureus in the chicken eggshell surface was 20.45% and 10.45%, respectively. Similarly, the prevalence of Staphylococcus spp. and S. aureus in quail eggshell surface was 16.25% and 5%, respectively. Overall, 27 isolates were identified as S. aureus, of which 23 were from the chicken eggshell surface and four from quail eggshell surface. Among the seven isolates tested, overall four (57.14%) were positive for the nuc gene. On the other hand, the mecA gene could be detected in three (50%) S. aureus out of six oxacillin resistant isolates. The antibiogram study indicated that most of the isolates were resistant to the antibiotics under ß-lactam group. CONCLUSION: The present study concludes that chicken and quail egg surface harbor multidrug-resistant bacteria which may cause public health hazards, if these antibiotic-resistant bacteria are transferred to a human.