Spatio-temporal dynamics and distributional trend analysis of African swine fever outbreaks (2020-2021) in North-East India.

African swine fever (ASF) is a highly contagious, notifiable, and fatal hemorrhagic viral disease affecting domestic and wild pigs. The disease was reported for the first time in India during 2020, resulted in serious outbreaks and economic loss in North-Eastern (NE) parts, since 47% of the Indian pig population is distributed in the NE region. The present study focused on analyzing the spatial autocorrelation, spatio-temporal patterns, and directional trend of the disease in NE India during 2020-2021. The ASF outbreak data (2020-2021) were collected from the offices of the Department of Animal Husbandry and Veterinary Services in seven NE states of India to identify the potential clusters, spatio-temporal aggregation, temporal distribution, disease spread, density maps, and risk zones. Between 2020 and 2021, a total of 321 ASF outbreaks were recorded, resulting in 59,377 deaths. The spatial pattern analysis of the outbreak data (2020-2021) revealed that ASF outbreaks were clustered in 2020 (z score = 2.20, p < .01) and 2021 (z score = 4.89, p < .01). Spatial autocorrelation and Moran's I value (0.05-0.06 in 2020 and 2021) revealed the spatial clustering and spatial relationship between the outbreaks. The hotspot analysis identified districts of Arunachal Pradesh, Assam and districts of Mizoram, Tripura as significant hotspots in 2020 and 2021, respectively. The spatial-scan statistics with a purely spatial and purely temporal analysis revealed six and one significant clusters, respectively. Retrospective unadjusted, temporal, and spatially adjusted space-time analysis detected five, five, and two statistically significant (p < .01) clusters, respectively. The directional trend analysis identified the direction of disease distribution as northeast-southwest (2020) and north-south (2021), indicate the possibility of ASF introduction to India from China. The high-risk zones and spatio-temporal pattern of ASF outbreaks identified in the present study can be used as a guide for deploying proper prevention, optimizing resource allocation and disease control measures in NE Indian states.

Pathotyping and antimicrobial susceptibility testing of Escherichia coli isolates from neonatal calves.

Neonatal calf mortality is a major concern to livestock sector worldwide. Neonatal calf diarrhoea (NCD), an acute severe condition causes morbidity and mortality in calves. Amongst various pathogens involved in NCD, E. coli is considered as one of the major causes. The study was targeted to characterize E. coli isolates from neonatal calves for diarrhoeagenic Escherichia coli (DEC) types (pathotyping), antimicrobial resistance (AMR) profiling and to correlate with epidemiological parameters. From neonates, a total of 113 faecal samples were collected, out of that 308, lactose fermenting colonies were confirmed as E. coli. Pathotypable isolates (12.3%) were represented by STEC (6.1%), EPEC (2.9%), ETEC (1.9%), EAEC (0.9%) and EHEC (0.3%). Occurrence of STEC was more in non-diarrhoeic calves, whereas ETEC was observed more in diarrhoeic calves. EPEC occurrence was observed in both diarrhoeic and non-diarrhoeic calves. Fishers extract test showed no significant association for occurrence of DEC types to type of dairies, health status, species, breed, age and sex of neonatal calves. Two hundred and eighty isolates were tested for antimicrobial susceptibility. The isolates showed maximum resistance towards ampicillin (55.4%) followed by tetracycline (54.3%), while minimum resistance was observed towards meropenem (2.5%). Multidrug resistant E. coli isolates were found to be 139 (49.6%), and Extended-spectrum beta-lactamase (ESBL) producers were 120 (42.9%). DEC pathotypes like STEC, ETEC, EHEC and EAEC that are also multidrug resistant present in neonatal calves have zoonotic potential and hence are of public health significance.

Multi-drug resistant (MDR), extended spectrum beta-lactamase (ESBL) producing and carbapenem resistant Escherichia coli in rescued Sloth bears (Melursus ursinus), India.

The study reports the multi-drug resistant (MDR), extended spectrum beta-lactamase (ESBL) producing and carbapenem resistant Escherichia coli (CRE) isolated from rescued sloth bear (Melursus ursinus), India. Non-duplicate faecal samples from 21 adult rescued sloth bears were collected at once during 2015-2016 and processed for isolation of E. coli and antibacterial susceptibility pattern. From 21 samples, 45 E. coli were isolated and on phenotypic screening, 23 were MDR, 17 were ESBL producers, and five were carbapenem-resistant (CR). Three E. coli isolates (6.67%, 3/45) showed no resistance, however 42 isolates (93.33%, 42/45) exhibited resistant to at least one antibiotics. The MDR isolates carried beta-lactamase, chloramphenicol, aminoglycosides, tetracycline, fluroquinolone, and sulphadimidine resistance genes. All the phenotypic ESBL producing isolates harbored blaCTX-M genes. On genotypic screening, three CRE (60.0%, 3/5) were positive for blaNDM carbapenemase gene and efflux pump-mediated carbapenem resistance was detected in two CRE isolates (40.0%, 2/5) which were negative for carbapenemase genes. The CRE isolates (n = 5) also co-harbored AMR genes like blaTEM-1, blaAmpC, qnrA, qnrB, qnrS, tetA, tetB and sulI. Virulence screening of the resistant isolates detected the presence of Stx1(n = 1), Stx2 (n = 3), eaeA (n = 4) and hlyA (n = 3) genes. Plasmid incompatibility (Inc) typing revealed that two isolates harboured blaNDM-5 gene on Incl1 and one isolate on IncF plasmid. Apart from the NDM gene, the plasmids also carried tetracycline, beta-lactamase and quinolone resistance genes. The plasmid multilocus sequence typing (pMLST) of the E. coli Incl1 plasmid showed the Sequence Type (ST) 297. This appears to be the first report of MDR, ESBL producing and blaNDM-5 genes on Incl1 and IncF plasmids from rescued sloth bear.

Faecal carriage of extended spectrum beta-lactamase (ESBL) and New Delhi metallo beta-lactamase(NDM) producing Escherichia coli between piglets and pig farmworkers.

A cross-sectional study on five organized pig farms was conducted to assess the faecal carriage of ESBL and blaNDM carbapenemase-producing E. coli in piglets and pig farmworkers. Faecal samples from piglets (n = 155) and pig farmworkers (n = 21) were processed for isolation and characterization of E. coli. A total of 124 E. coli isolates from piglets and 21 E. coli isolates pig farmworkers were recovered and screening for ESBL production showed that 44.4 % (55/124) of the isolates from piglets and 42.9 % (9/21) of the isolates from farmworkers were ESBL positive. The ESBL positive isolates from piglets and farmworkers harbored blaCTX-M and also co-harbored other beta-lactams, sulphonamide, quinolone and tetracycline resistance genes. Diarrhoeic (50%, 49/98) and crossbred piglets (52.7%, 39/74) harbored a significantly higher number of ESBL producing isolates than non-diarrhoeic (23.1 %, 6/26) and purebred piglets (32%, 16/50) (p < 0.05). Piglets and pig farmworkers harbored nine and two carbapenem-resistant isolates, respectively. Interestingly, two isolates from piglets and one isolate from farmworkers harbored the blaNDM gene. The blaNDM positive E. coli isolated from piglets and farmworkers of the same farm revealed similar antibacterial resistance patterns, resistant genes, sequence (ST-167) and plasmid type (IncX3). In India, carbapenems are not used in food animal treatment, hence carbapenem resistant E. coli in piglets possibly originated from the human contact or common environment and is of public health importance.

Bluetongue in India: a systematic review and meta-analysis with emphasis on diagnosis and seroprevalence.

Bluetongue (BT) is an infectious viral disease which affects a wide range of ruminants and was first reported in India in 1964. In view of the absence of comprehensive information on the BT status in India, this study presents the seroprevalence on BT in farm animals of India based-on a systematic review and meta-analysis. A systematic review was conducted to identify the published articles (2001-2018) reporting the seroprevalence of BT in sheep, goats, cattle, buffalo, camels, and Mithun (Bos frontalis) from India. From 409 research articles, 71 fulfilled the inclusion criteria and meta-analysis for proportions was carried out targeting the eligible studies. From these, 144 strata level data were extracted with a sample size of 14048 sheep, 14696 goats, 5218 cattle, 2653 buffaloes, 2062 camels, and 222 Mithun. Overall, the analyses showed that the BT seroprevalence of 43% (95% CI: 38-49%) in goats, 39% (95% CI: 33-46%) in sheep, 38% (95% CI: 25-45%) in cattle, 34% (95% CI: 20-51%) in buffaloes, 16% (95% CI: 10-22%) in camels, and 66% (95% CI: 17-95%) in Mithun. Furthermore, the meta-regression analysis suggested that serological tests, geographical region, and sample size were the prime moderators. Meta-analytic study indicates the BT seropositivity in 25.35 million sheep (95% CI: 21.5-29.9), 58 million goats (95% CI: 51.3-66.2), 66.8 million cattle (95% CI: 47.7-86), 37.0 million buffaloes (95% CI: 21.7-55.4), 0.06 million camels (95% CI: 0.04-0.09), and 0.19 million Mithun (95% CI: 0.05-0.28). The findings highlight the variation of BT seropositivity in different geographical regions of India.

Rhinosporidiosis in Central India: A cross-sectional study from a tertiary care hospital in Chhattisgarh.

BACKGROUND AND AIMS: Chhattisgarh is an endemic region for rhinosporidiosis infection in Central India. This high prevalence can be explained by suitable hot tropical environment as well as social practice of common bathing of public with animals outdoor, mainly in ponds and river. There are more than 200 ponds in and around Bilaspur district, mainly in Ratanpur region. However, due to limited literature describing rhinosporidiosis from this part of India, it remains neglected and still not recognized as a serious health-care issue and any definite government or private measures/initiatives are not taken to counter this problem. MATERIALS AND METHODS: It is a retrospective demographic study analyzing 113 consecutive biopsies received from different clinical departments in a tertiary care center with a histological diagnosis of rhinosporidiosis over a period of 7 years from January 2011 to December 2017. RESULTS: Most of the patients presented in the otorhinolaryngology (86.7%) department, followed by ophthalmology (10.6%). The mean age of the patients was 23.4 years, with a range of 3-70 years. A male-to-female ratio of about 3:2 was noted. Nasal cavity was the most common site of involvement (90/113), followed by conjunctiva (7/113), nasal cavity with oropharynx (6/113), and eyelid (3/113). Patients' complaints showed a varying range from nasal block, nasal mass, bleeding, bleed on touch, difficulty in breathing, and conjunctival mass. Two of the cases presented as recurrent rhinosporidiosis. CONCLUSION: Rhinosporidiosis is endemic and possesses a great health-care burden in Chhattisgarh. The socioeconomic status, outdoor bathing, and cultural practice in rural areas are the major factors contributing to newer cases. An urgent government initiative to decontaminate public water bodies and public health education through health-care workers and nongovernment organizations is the need of the hour to curtail the incidence of rhinosporidiosis.

Cross-sectional study on rotavirus A (RVA) infection and assessment of risk factors in pre- and post-weaning piglets in India.

Rotavirus A (RVA) infections are known to retard the piglets' growth and minimize the profit to the pig farming community. Between August 2014 and July 2017, in a cross-sectional study, we surveyed 13 organized pig farms located in the eight states of India representing northern, north-eastern and southern regions, to identify the risk factors associated with RVA infection in pre- and post-weaning piglets. Faecal samples (n = 411) comprising of non-diarrhoeic (n = 320) and diarrhoeic (n = 91) were collected and screened for RVA infection using VP6 gene-based RT-PCR. RVA positivity of 52.5% (168/320) in non-diarrhoeic and 59.3% (54/91) in diarrhoeic piglets was noticed. Further, 53.3% (120/225) and 54.8% (102/186) of the samples from pre- and post-weaned samples were positive for RVA, respectively. To note, no statistically significant association was noticed between RVA infection, health and weaning status. Additionally, a questionnaire-based survey was conducted to identify the risk factors for RVA infections in piglets. The analysis revealed that good ventilation (OR 0.2, 95% CI 0.15-0.39), use of deep well water (OR 0.2, 95% CI 0.13-0.43) and feeding of commercial feed (OR 0.3, 95% CI 0.18-0.41) were associated with reduced risk of RVA infection compared with poor ventilation, use of shallow well water and feeding of own milled feed, respectively. Contrarily, mixed farms (OR 2.1, 95% CI 1.26-3.37), use of heater or cooler (OR 5.9, 95% CI 3.74-9.30), sheds in different elevation (OR 2.5, 95% CI 1.20-5.01) and weekly and occasional use of disinfectant for surface cleaning (OR 1.8, 95% CI 1.12-2.96) were associated with higher RVA infection. Mitigating the risk factors might help in better health management of piglets and increase the economic return to pig farming community in the country.

Impact of HfO2 as a Passivation Layer in the Solar Cell Efficiency Enhancement in Passivated Emitter Rear Cell Type.

We report the simulation of high-efficiency c-silicon Passivated Emitter Rear Contact (PERC) type solar cell structure with rear side passivated with HfO2 as a passivating material. Variation in the half length of pyramid has been carried out to investigate its effect on the solar cell electrical characteristics such as fill factor (FF), open circuit voltage (Voc) and efficiency. Aluminum back Surface Field (Al-BSF) and PERC type solar cell with Al2O3 passivation layer structures were also modeled for comparison. Effect of variation in passivation layer (HfO2) thickness (10 and 15 nm) and permittivity (k = 14 and 25) on the solar cell electrical characteristics has been investigated. Result shows the efficiency improvement in the PERC solar cell with HfO2 passivation layer by 0.5941% and 0.983% as compared to the Al-BSF and PERC with Al2O3 passivation layer at 8 µm pyramid half length. Increased series resistance and reduced FF has been observed with the incorporation of passivation layer at the solar cell structure. Negligible effect of passivation layer thickness has been observed on the solar cell electrical parameters whereas the permittivity value does have significant effect.

Risk factor analysis, antimicrobial resistance and pathotyping of Escherichia coli associated with pre- and post-weaning piglet diarrhoea in organised farms, India.

A cross-sectional study was conducted from 2014 to 2017 in 13 organised pig farms located in eight states of India (Northern, North-Eastern and Southern regions) to identify the risk factors, pathotype and antimicrobial resistance of Escherichia coli associated with pre- and post-weaning piglet diarrhoea. The data collected through questionnaire survey were used to identify the risk factors by univariable analysis, in which weaning status, season, altitude, ventilation in the shed, use of heater/cooler for temperature control in the sheds, feed type, water source, and use of disinfectant, were the potential risk factors. In logistic regression model, weaning and source of water were the significant risk factors. The piglet diarrhoea prevalence was almost similar across the regions. Of the 909 faecal samples collected (North - 310, North-East - 194 and South - 405) for isolation of E. coli, pathotyping and antibiotic screening, 531 E. coli were isolated in MacConkey agar added with cefotaxime, where 345 isolates were extended spectrum ß-lactamase (ESBL) producers and were positive for blaCTX-M-1 (n = 147), bla TEM (n = 151), qnrA (n = 98), qnrB (n = 116), qnrS (n = 53), tetA (n = 46), tetB (n = 48) and sul1 (n = 54) genes. Multiple antibiotic resistance (MAR) index revealed that 14 (2.64%) isolates had MAR index of 1. On the virulence screening of E. coli, 174 isolates harboured alone or combination of Stx1, Stx2, eaeA, hlyA genes. The isolates from diarrhoeic and post-weaning samples harboured higher number of virulence genes than non-diarrhoeic and pre-weaning. Alleviating the risk factors might reduce the piglet diarrhoea cases. The presence of multidrug-resistant and ESBL-producing pathogenic E. coli in piglets appears a public health concern.

Epidemiology of carbapenem-resistant Escherichia coli and first report of blaVIM carbapenemases gene in calves from India.

A cross-sectional study on six dairy farms was conducted to ascertain the occurrence of carbapenem-resistant Escherichia coli in calves. Two-hundred and seventy-nine isolates of E. coli were recovered from 90 faecal samples from apparently healthy (45) and diarrhoeal (45) calves. The isolates were screened for phenotypic susceptibility to carbapenems and production of metallo ß-lactamase, as well as five carbapenemase resistance genes by PCR, and overexpression of efflux pumps. Eighty-one isolates (29.03%) were resistant to at least one of three carbapenem antibiotics [meropenem (23.30%), imipenem (2.15%) and ertapenem (1.43%)], and one isolate was positive for the blaVIM gene which was located on an Incl1 plasmid of a novel sequence type (ST 297) by multilocus sequence typing. The majority (83.95%) of isolates had an active efflux pump. Calves housed on concrete floors were approximately seven times more likely to acquire meropenem-resistant isolates than those housed on earthen floors (95% CI 1.27-41.54). In India, carbapenem drugs are not used in food animal treatment, hence carbapenem-resistant strains in calves possibly originate from the natural environment or human contact and is of public health importance. To our knowledge, this is the first report of blaVIM carbapenemases gene in calves from India.

Design and fabrication of a magnetically actuated non-invasive reusable drug delivery device.

We present a novel approach of designing and fabricating a noninvasive drug delivery device which is capable of delivering the drug to the target site in a controlled manner. The device utilizes a reservoir which can be reused once the drug has completely diffused from it. This micro-reservoir based fabricated device has been successfully tested using niosomes of insulin drug filled in, which was then sealed with a magnetic membrane of 20 µm thick and was actuated by applying magnetic field. The deflection of the membrane on application of magnetic field results in the drug release from the reservoir. The discharge of the drug solution and the release rates was controlled by external magnetic field. The simulation of the membrane deflection using COMSOL software was carried out to optimize the concentration of the ferrous nanopowder in PDMS matrix. The characterization of the devices was implemented in-vitro on water and in-vivo on Wistar rats. It was also validated using high-performance liquid chromatography (HPLC) by observing characteristic peak of insulin. The blood samples showed the retention time of 2.79 min at λmax of 280 nm which further authenticated the effectiveness of the proposed work. This noninvasive fabricated device provides reusability, precise control and can enable the patient or a physician to actively administrate the drug when required.

Molecular characterisation of blaOXA-48 carbapenemase-, extended-spectrum ß-lactamase- and Shiga toxin-producing Escherichia coli isolated from farm piglets in India.

OBJECTIVES: The aim of this study was to characterise carbapenemase-, extended-spectrum ß-lactamase (ESBL)- and Shiga toxin-producing Escherichia coli isolated from farm piglets in India. METHODS: Faecal samples (n=741) from 10 organised pig farms, including non-diarrhoeic (n=546) and diarrhoeic (n=195) piglets, were processed for isolation of carbapenem-resistant and ESBL-producing E. coli. RESULTS: A total of 27 and 243 isolates were phenotypically confirmed as carbapenem-resistant and ESBL-producers, respectively. The meropenem minimum inhibitory concentration (MIC) of carbapenem-resistant isolates ranged from 8-128µg/mL. On genotypic screening of the 27 carbapenem-resistant isolates, 3 isolates were positive for the blaOXA-48 carbapenemase gene; no other carbapenemase genes were detected. The 243 ESBL-producing isolates were positive for blaCTX-M-1 (n=135), qnrA (n=92), qnrB (n=112), qnrS (n=49), tetA (n=42), tetB (n=45) and sul1 (n=43). The Shiga toxin virulence markers stx1 and stx2 were detected in 41 and 38 of the 243 phenotypic ESBL-producing isolates, respectively. Multilocus sequence typing (MLST) of blaOXA-48-positive E. coli isolates showed ST10- and ST5053-like sequence types. CONCLUSION: This is the first report on the presence of blaOXA-48-carrying E. coli in piglets in India, which pose a potential risk to public health.

Spatial molecular epidemiology of carbapenem-resistant and New Delhi metallo beta-lactamase (blaNDM)-producing Escherichia coli in the piglets of organized farms in India.

AIM: A cross-sectional study was conducted in 10 government-organized pig farms between 2014 and 2016 representing seven states of India to understand the epidemiology of carbapenem resistance in the Escherichia coli. METHODS AND RESULTS: In this study, fecal sample (n = 673) from non-diarrheic (n = 501) and diarrheic (n = 172) piglets were processed for isolation of carbapenem resistant E. coli. Of 673, E. coli isolate (n = 112) was genotyped for confirming the carbapenem resistance and associated virulence factors. Of the 112 isolates, 23 were phenotypically resistant to carbapenem and 8 were carrying the New Delhi metallo beta-lactamase (blaNDM) gene. The carbapenem-resistant isolates also produced extended spectrum beta-lactamases and were multidrug resistant. The PCR-based pathotyping revealed the presence of stx1, stx2, eae and hlyA genes. The enterobacterial repetitive intergenic consensus PCR dendrogram analysis of the isolates yielded three distinct clusters. The statistical analysis revealed no association between carriages of carbapenem-resistant E. coli in different breed of piglets however, location, sex, health status of piglets and age showed significant difference. The spatial analysis with SaTScan helped in identification of carbapenem-resistant clusters. CONCLUSIONS: The presence of carbapenem resistant E. coli isolates with virulence genes in the piglet poses a potential public health risk through possible access and spread via the food chain and environment. Efflux pump may also play an important role in carbapenem resistance in piglet E. coli isolates. Furthermore, identification of risk factors in relation to spatial clusters will help in designing preventive strategies for reducing the risk of spread of carbapenem resistant bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: 1. Piglets harbor carbapenem resistant E. coli and have great public health significance. 2. Apart from carbapenemase, efflux pump is also important for carbapenem resistance. 3. This is the first report of blaNDM in the piglets from India.

Anaerobic digestion of sewage sludge with grease trap sludge and municipal solid waste as co-substrates.

The feasibility of simultaneous treatment of multiple wastes via co-digestion was studied in semi-continuous mode at mesophilic conditions. The obtained results indicated that sewage sludge, organic fraction of municipal waste (OFMSW) and grease trap sludge (GTS) possess complementary properties that can be combined for successful anaerobic digestion. During the co-digestion period, methane yield and VS removal were significantly higher in comparison to digestion of sewage sludge alone. Addition of GTS to digesters treating sewage sludge resulted in increased VS removal and methane yield up to 13% (from 50 to 56.4) and 52% (from 300 to 456,547m3/Mg VSadd), respectively. While the use of OFMSW as the next co-substrate in the feedstock, can boost methane yield and VS removal up to 82% (300-547m3/Mg VSadd) and approximately 29% (from 50% to 64.7%), respectively. Moreover, the results of the present laboratory study revealed that the addition of co-substrates to the feedstock had a significant influence on biogas composition. During the experiment methane content in biogas ranged from 67% to 69%. While, the concentration of LCFAs was increasing with the gradual increase in the share of co-substrates in the mixtures, wherein only the oleic acid was higher than some inhibition concentrations which have been reported in the literature. However, it did not significantly affect the efficiency of the co-digestion process.

Black tea: Phytochemicals, cancer chemoprevention, and clinical studies.

Tea (Camellia sinensis L.) is the most popular, flavored, functional, and therapeutic non-alcoholic drink consumed by two-thirds of the world's population. Black tea leaves are reported to contain thousands of bioactive constituents such as polyphenols, amino acids, volatile compounds, and alkaloids that exhibit a range of promising pharmacological properties. Due to strong antioxidant property, black tea inhibits the development of various cancers by regulating oxidative damage of biomolecules, endogenous antioxidants, and pathways of mutagen and transcription of antioxidant gene pool. Regular drinking of phytochemicals-rich black tea is linked to regulate several molecular targets, including COX-2, 5-LOX, AP-1, JNK, STAT, EGFR, AKT, Bcl2, NF-κB, Bcl-xL, caspases, p53, FOXO1, TNFα, PARP, and MAPK, which may be the basis of how dose of black tea prevents and cures cancer. In vitro and preclinical studies support the anti-cancer activity of black tea; however, its effect in human trails is uncertain, although more clinical experiments are needed at molecular levels to understand its anti-cancer property. This review discusses the current knowledge on phytochemistry, chemopreventive activity, and clinical applications of black tea to reveal its anti-cancer effect.

Selenium and other mineral concentrations in feed and sheep's blood in Kosovo.

The aim of this study was to assess the concentration of Se and other minerals in sheep and the supplied feed. Four macrominerals (Ca, P, Mg, and S), 7 microminerals (Se, Fe, Zn, Cu, Mn, Co, and Mo), and 2 toxic minerals (Cd and Pb) were analyzed in 69 feed and 292 sheep blood samples from 30 farms in different regions of Kosovo. The samples were analyzed using inductively coupled plasma mass spectrometry, and mineral concentrations in whole blood were measured to assess their status in animals. Concentrations of the different minerals in feed were found in the following ranges: 1.9 to 9.5 g Ca/kg DM, 0.8 to 3.2 g P/kg DM, 0.8 to 3.2 g Mg/kg DM, 1.0 to 2.8 g S/kg DM, 6 to 82 µg Se/kg DM, 33 to 970 mg Fe/kg DM, 15 to 42 mg Zn/ kg DM, 2.6 to 7.5 mg Cu/kg DM, 26 to 250 mg Mn/kg DM, 0.04 to 0.88 mg Co/kg DM, 0.05 to 0.86 mg Mo/ kg DM, 0.07 to 2.02 mg Pb/kg DM, and 0.02 to 0.19 mg Cd/kg DM. Concentrations of the microminerals analyzed in whole blood were found in the following ranges: 15 to 360 µg Se/L, 190 to 500 mg Fe/L, 1.4 to 3.8 mg Zn/L, 0.3 to 2.6 mg Cu/L, 6 to 243 µg Mn/L, 0.1 to 19.6 µg Co/L, and 1.8 to 66.0 µg Pb/L. Among all minerals, the largest deficiency was found for Se both in feed and sheep blood, with 82% of feed samples and 83% blood samples being inadequate, and its supplementation is necessary. Selenium-supplemented sheep had significantly higher Se concentration in blood than non-supplemented sheep (P < 0.01). In addition, other macro- and microminerals in feed such as P, S, Cu, and Co were at inadequate concentrations at some of the farms, and supplementation may also be needed for these minerals.

Quercetin sensitizes fluconazole-resistant candida albicans to induce apoptotic cell death by modulating quorum sensing.

Quorum sensing (QS) regulates group behaviors of Candida albicans such as biofilm, hyphal growth, and virulence factors. The sesquiterpene alcohol farnesol, a QS molecule produced by C. albicans, is known to regulate the expression of virulence weapons of this fungus. Fluconazole (FCZ) is a broad-spectrum antifungal drug that is used for the treatment of C. albicans infections. While FCZ can be cytotoxic at high concentrations, our results show that at much lower concentrations, quercetin (QC), a dietary flavonoid isolated from an edible lichen (Usnea longissima), can be implemented as a sensitizing agent for FCZ-resistant C. albicans NBC099, enhancing the efficacy of FCZ. QC enhanced FCZ-mediated cell killing of NBC099 and also induced cell death. These experiments indicated that the combined application of both drugs was FCZ dose dependent rather than QC dose dependent. In addition, we found that QC strongly suppressed the production of virulence weapons-biofilm formation, hyphal development, phospholipase, proteinase, esterase, and hemolytic activity. Treatment with QC also increased FCZ-mediated cell death in NBC099 biofilms. Interestingly, we also found that QC enhances the anticandidal activity of FCZ by inducing apoptotic cell death. We have also established that this sensitization is reliant on the farnesol response generated by QC. Molecular docking studies also support this conclusion and suggest that QC can form hydrogen bonds with Gln969, Thr1105, Ser1108, Arg1109, Asn1110, and Gly1061 in the ATP binding pocket of adenylate cyclase. Thus, this QS-mediated combined sensitizer (QC)-anticandidal agent (FCZ) strategy may be a novel way to enhance the efficacy of FCZ-based therapy of C. albicans infections.

Colostrum hexasaccharide, a novel Staphylococcus aureus quorum-sensing inhibitor.

The discovery of quorum-sensing (QS) systems regulating antibiotic resistance and virulence factors (VFs) has afforded a novel opportunity to prevent bacterial pathogenicity. Dietary molecules have been demonstrated to attenuate QS circuits of bacteria. But, to our knowledge, no study exploring the potential of colostrum hexasaccharide (CHS) in regulating QS systems has been published. In this study, we analyzed CHS for inhibiting QS signaling in Staphylococcus aureus. We isolated and characterized CHS from mare colostrum by high-performance thin-layer chromatography (HPTLC), reverse-phase high-performance liquid chromatography evaporative light-scattering detection (RP-HPLC-ELSD), (1)H and (13)C nuclear magnetic resonance (NMR), and electrospray ionization mass spectrometry (ESI-MS). Antibiofilm activity of CHS against S. aureus and its possible interference with bacterial QS systems were determined. The inhibition and eradication potentials of the biofilms were studied by microscopic analyses and quantified by 96-well-microtiter-plate assays. Also, the ability of CHS to interfere in bacterial QS by degrading acyl-homoserine lactones (AHLs), one of the most studied signal molecules for Gram-negative bacteria, was evaluated. The results revealed that CHS exhibited promising inhibitory activities against QS-regulated secretion of VFs, including spreading ability, hemolysis, protease, and lipase activities, when applied at a rate of 5 mg/ml. The results of biofilm experiments indicated that CHS is a strong inhibitor of biofilm formation and also has the ability to eradicate it. The potential of CHS to interfere with bacterial QS systems was also examined by degradation of AHLs. Furthermore, it was documented that CHS decreased antibiotic resistance in S. aureus. The results thus give a lead that mare colostrum can be a promising source for isolating a next-generation antibacterial.

Trichoderma harzianum elicits induced resistance in sunflower challenged by Rhizoctonia solani.

AIMS: To investigate the efficacy of Trichoderma harzianum NBRI-1055 (denoted as 'T-1055') in suppression of seedling blight of sunflower caused by Rhizoctonia solani Kühn and their impact on host defence responses. METHODS AND RESULTS: T-1055 was applied as seed treatment, soil application and combined application (seed treatment + soil application). Higher protection afforded by combined application of T-1055 was associated with the marked induction of phenylalanine ammonia-lyase (PAL), polyphenol oxidase (PPO), peroxidase (PO) and cinnamyl alcohol dehydrogenase (CAD) activities. The activities of PAL and PPO reached maximum at 10 days after sowing (DAS), while PO and CAD levels reached maximum at 12 DAS. This was further supported by the accumulation of total phenolic content that showed an increase up to threefold at 14 DAS. In addition, HPLC analysis revealed that the contents of ferulic and p-coumaric acids increased by 6·3 and 4·6 times, respectively, at 14 DAS. Amount of gallic acid was also little more than double. Lignin deposition in sunflower root increased by 2·7, 3·4 and 3·7 times through combined application of T-1055 at 16, 18 and 20 DAS, respectively. Combined application also increased the accumulation of PR-2 and PR-3 proteins by 3·3 and 3·9 times, respectively, at 12 DAS in followed by seed treatment alone. CONCLUSIONS: The combined application of T-1055 triggered defence responses in an enhanced level in sunflower than the soil and seed alone and provided better protection against Rhizoctonia seedling blight. SIGNIFICANCE AND IMPACT OF THE STUDY: Rhizospheric fungal bioagent 'T-1055' can enhance protection in sunflower against the R. solani pathogen through augmented elicitation of host defence responses.