Functional divergence of white genes in Henosepilachna vigintioctopunctata revealed by RNA interference.

Henosepilachna vigintioctopunctata is a serious pest of Solanaceae and Cucurbitaceae in many Asian countries. RNA interference (RNAi) can effectively reduce transcript abundance in this beetle, offering opportunities to explore the biological function of specific genes. The white gene encodes a half-type ATP-binding cassette transporter that plays an essential role in tryptophan, guanine and uric acid transport across membranes. Mutations that disrupt the function of white are known to cause eye pigmentation phenotypes in many insect species. Here, we found evidence for five white gene paralogues present in H. vigintioctopunctata transcriptome datasets sequenced from a range of developmental stages. We individually knocked down each of the five white genes through the injection of corresponding double-stranded RNAs (dsRNAs) to the fourth-instar larvae to determine whether functional divergence has occurred. We found that injecting 1 µg dswhite3 caused compound eye colour of pupae and adults to develop as red/brown and brown, respectively, compared with black eyes in control beetles. Injection of 2 µg dswhite3 increased RNAi efficacy and produced a clearer eye colour phenotype. At both doses, the ocular diaphragm (a ring of black pigment surrounding each eye) did not change in the white3 RNAi hypomorphs. Moreover, our data revealed that injection of dswhite2 at the fourth-instar larval stage impaired the climbing ability of both male and female adults. Our results confirmed, for the first time, functional divergence of duplicated white genes in an insect species.

Prevalence of Degnala disease in bovine along with screening of toxigenic fungi isolated from contaminated rice straw.

Toxigenic potential of different candidate fungi, isolated from rice straw feed of Degnala disease affected bovines was analyzed along with species, age, gender and seasonal prevalence. Of 1,536 cases, 104 (6.77%) showed positive signs with a significant association (p less than 0.05) between this disease and rice straw feeding, in buffaloes, and bovine aged over 1 year in the winter season. Complete blood count showed a marked increase in erythrocyte sedimentation rate and all white blood cells numbers, except lymphocytes in positive cases. There was a significant increase (p less than 0.05) in alanine amino transferase, aspirate amino transferase and alkaline phosphatase in the liver function test. At the same time, an increased value of creatinine was noted in the renal function test. For isolation and screening of toxigenic fungi, rice straw samples (n=40) being fed to the positive cases were processed further, and 85 fungal isolates were found, mainly of Aspergillus (57), Penicillium (10), Fusarium (04), Zygomycetes (03), Curvularia (01) and unidentified (10). All isolated fungi were subjected for mycotoxin production and only 11 showed mycotoxin-producing capability (including Aspergillus, Penicillium and Fusarium isolates) analyzed by thin layer chromatography and quantified through high performance liquid chromatography. It is concluded that all the fungi contaminating rice straw feed of Degnala affected animals were not toxigenic. This work will help in establishing major mycotoxin-producing fungi leading to the probable cause of Degnala disease in bovine.

Outcomes after endovascular or open repair for degenerative descending thoracic aortic aneurysm using linked hospital data.

BACKGROUND: The population-based effectiveness of thoracic endovascular aortic repair (TEVAR) versus open surgery for descending thoracic aortic aneurysm remains in doubt. METHODS: Patients aged over 50 years, without a history of aortic dissection, undergoing repair of a thoracic aortic aneurysm between 2006 and 2011 were assessed using mortality-linked individual patient data from Hospital Episode Statistics (England). The principal outcomes were 30-day operative mortality, long-term survival (5 years) and aortic-related reinterventions. TEVAR and open repair were compared using crude and multivariable models that adjusted for age and sex. RESULTS: Overall, 759 patients underwent thoracic aortic aneurysm repair, mainly for intact aneurysms (618, 81·4 per cent). Median ages of TEVAR and open cohorts were 73 and 71 years respectively (P < 0·001), with more men undergoing TEVAR (P = 0·004). For intact aneurysms, the operative mortality rate was similar for TEVAR and open repair (6·5 versus 7·6 per cent; odds ratio 0·79, 95 per cent confidence interval (c.i.) 0·41 to 1·49), but the 5-year survival rate was significantly worse after TEVAR (54·2 versus 65·6 per cent; adjusted hazard ratio 1·45, 95 per cent c.i. 1·08 to 1·94). After 5 years, aortic-related mortality was similar in the two groups, but cardiopulmonary mortality was higher after TEVAR. TEVAR was associated with more aortic-related reinterventions (23·1 versus 14·3 per cent; adjusted HR 1·70, 95 per cent c.i. 1·11 to 2·60). There were 141 procedures for ruptured thoracic aneurysm (97 TEVAR, 44 open), with TEVAR showing no significant advantage in terms of operative mortality. CONCLUSION: In England, operative mortality for degenerative descending thoracic aneurysm was similar after either TEVAR or open repair. Patients who had TEVAR appeared to have a higher reintervention rate and worse long-term survival, possibly owing to cardiopulmonary morbidity and other selection bias.

Prevalence of Gingivitis, Plaque accumulation and Decayed, Missing and Filled Teeth among slum population in Bangladesh.

A cross-sectional survey, using cluster sampling technique, of slum population, was done to explore the oral health status and the prevalence of common oral diseases. A close ended questionnaire comprising Decayed, Missing and Filled Teeth (DMFT) Index, Gingival Index (Löe and Silness) and Plaque Index was applied to evaluate and record oral diseases, in both male and female population, covering a wide range of age groups. Clinical examination was carried out in different shum set ups, including slum schools by trained and calibrated examiners. Three thousand nine hundred and four (3904) slum dwellers participated in the survey. Prevalence of Caries was expressed in mean DMFT, recording of gingival status followed the method of Löe and Silness, oral hygiene status was evaluated using Plaque index. Mean decayed component, of the DMFT, was significantly higher than filling and missing component. Both decayed and missing components showed increasing trend, and filling components decreased as the age progressed. Prevalence of gingivitis and plaque accumulation was remarkably high among slum dwellers. Significantly high level of common oral diseases was found among Tongi slum dwellers.

Technology platform development for targeted plasma metabolites in human heart failure.

BACKGROUND: Heart failure is a multifactorial disease associated with staggeringly high morbidity and motility. Recently, alterations of multiple metabolites have been implicated in heart failure; however, the lack of an effective technology platform to assess these metabolites has limited our understanding on how they contribute to this disease phenotype. We have successfully developed a new workflow combining specific sample preparation with tandem mass spectrometry that enables us to extract most of the targeted metabolites. 19 metabolites were chosen ascribing to their biological relevance to heart failure, including extracellular matrix remodeling, inflammation, insulin resistance, renal dysfunction, and cardioprotection against ischemic injury. RESULTS: In this report, we systematically engineered, optimized and refined a protocol applicable to human plasma samples; this study contributes to the methodology development with respect to deproteinization, incubation, reconstitution, and detection with mass spectrometry. The deproteinization step was optimized with 20% methanol/ethanol at a plasma:solvent ratio of 1:3. Subsequently, an incubation step was implemented which remarkably enhanced the metabolite signals and the number of metabolite peaks detected by mass spectrometry in both positive and negative modes. With respect to the step of reconstitution, 0.1% formic acid was designated as the reconstitution solvent vs. 6.5 mM ammonium bicarbonate, based on the comparable number of metabolite peaks detected in both solvents, and yet the signal detected in the former was higher. By adapting this finalized protocol, we were able to retrieve 13 out of 19 targeted metabolites from human plasma. CONCLUSIONS: We have successfully devised a simple albeit effective workflow for the targeted plasma metabolites relevant to human heart failure. This will be employed in tandem with high throughput liquid chromatography mass spectrometry platform to validate and characterize these potential metabolic biomarkers for diagnostic and therapeutic development of heart failure patients.

Incidence of descending aortic pathology and evaluation of the impact of thoracic endovascular aortic repair: a population-based study in England and Wales from 1999 to 2010.

OBJECTIVES: To investigate population trends in thoracic aortic disease (dissections and aneurysms) in England and Wales, with focus on the impact of thoracic endovascular aortic repair on procedure numbers and age at repair. MATERIALS AND METHODS: Routine hospital statistics of England and Wales provided admission, procedure and mortality data from 1999 to 2010. All data were age-standardised, reported per 100,000 population, by age bands (>50 years or 50-74 years versus 75+ years) and gender. Only patients 50+ years were included, to focus on degenerative disease. RESULTS: Between 1999 and 2010 hospital admissions for total (ascending and descending) have risen steadily for thoracic aortic dissection (TAD) from 7.2 to 8.8 and thoracic aortic aneurysm (TAA) from 4.4 to 9.0, principally attributable to increased admissions in those 75+ years. Total mortality declined steadily over the same period, for TAD from 4.4 to 3.2 and for TAA from 10.4 to 7.5. Procedure rates have risen sharply, driven by the implementation of TEVAR from 2006, for type B dissection from 0.06 to 0.53 and for descending TAA from 0.76 to 1.89. All figures are per 100,000 population with P <0.005. CONCLUSION: Improvements in case ascertainment may have contributed to the increase in hospital admissions. The increased application of TEVAR, particularly for dissections, is mainly in those above 75 years and has not yet translated into an accelerated survival benefit.

Immuno-affinity chromatography for purification of IgG from hyper-immune sera raised against 146S fraction of Foot and Mouth Disease Virus for diagnostic purposes.

Immunoaffinity chromatography (IAC) is a fundamental isolation and purification tool which is incorporated in a substantial range of therapeutic and diagnostic applications. This study has reappraised the usefulness of immunoaffinity chromatography for the purification of polyclonal antibodies. Protein A based IAC is a convenient and reliable method for purification of IgG, from hyperimmunesera (HIS) raised in experimental animals such as rabbits, guinea pigs and mice to be utilized in pharmaceutics and diagnostics. The 146S fraction of Foot and Mouth Disease virus (FMDV) TCID50=10 5.6 was cultured on a baby hamster kidney cell line 21 (BHK-21), concentrated using salt precipitation method using PEG 6000, purified by size exclusion chromatography (SEC) using Sepharose-30 at 254nm absorbance. Purification of 146S FMDV was analyzed using 12% SDS-PAGE which provided two bands of light and heavy chains. The alum-based vaccine, consisting of ≥10µg of 146S FMDV, was applied in 10 male rabbits and 10 male guinea pigs and two animals of each group were taken as a negative control. The titer of serum was calculated using virus neutralization test. A Protein-A kit (Thermo scientific- 44667, 0528.2) was used to purify HIS raised against 146S FMDV and validated using 12% SDS PAGE in reducing condition. The data demonstrate that protein-A affinity chromatography is an efficient tool for the purification of antibodies from hyper-immune sera raised against 146S FMDV and can be used for the production of diagnostic kits e.g. Enzyme linked immuno-sorbent assay (ELISA) and radioimmunoassay.

Evolutionary dynamics of topotype ME-SA/Ind-2001 of foot and mouth disease virus serotype-O in Pakistan: 2017-2022.

Background: During past few years, the Ind-2001 lineage of the Middle East-South Asia topotype (ME-SA) of the foot-and-mouth disease (FMD) virus has been implicated in FMD outbreaks in Pakistan. Aims: This work conducts a comprehensive evolutionary analysis of the Ind-2001 and Pan Asia II lineages, with a specific emphasis on their geographical distribution, lineage classification, and sub-lineage distribution within the region. Furthermore, it aims to expand our understanding of the conserved region of the VP1 protein. Methods: Total samples (n=50) were subjected to antigen detection ELISA and RT-PCR for serotype determination. Confirmed serotype-O isolates (n=17) underwent sequencing for lineage comparison, mutation impact assessment on the VP1 protein GH loop, 3D structure prediction, and further comparative analysis. Results: Isolates collected from 2017 to 2020 were identified as serotypes O/ME-SA/Pan Asia II ANT10 and O/ME-SA/Pak14. Notably, isolates collected from 2020 to 2022 belonged to a novel FMDV serotype O/ME-SA/Ind-2001e lineage. Phylogenetic analyses indicated that these strains were distinct from dominant contemporaneous strains which may challenge Pakistan's FMD control measures. These isolates exhibited variance in the VP1 epitope, specifically in amino acid residues 135-155, known to influence neutralizing antibody generation. Conclusion: Observed mutations suggest potential challenges to current vaccination efficacy against FMD. This emphasizes enhanced FMD surveillance and demonstrates that tracking the emergence of the O/ME-SA/Ind-2001e lineage is important for determining FMD control strategies in Asia.

Biofilm forming multidrug resistant Staphylococcus aureus of dairy origin: molecular and evolutionary perspectives.

Background: Biofilm production by Staphylococcus aureus is a prevailing cause of multidrug resistance. The evolutionary mechanisms of adaption with host and pathogenicity are poorly understood. Aims: The present study aimed to investigate the biofilm-forming potential, associated multidrug resistance, and the evolutionary analysis of S. aureus isolated from bovine subclinical mastitis. Methods: 122 S. aureus isolates were subjected to Congo red agar method (CRA), microtitre plate method (MTP), and PCR to check the biofilm-forming potential. The Kirby-Bauer disk diffusion method was used to evaluate the antibiotic resistance pattern. The icaA gene of isolates was subjected to molecular and evolutionary analysis using different bioinformatics tools. Results: The results showed that 63.93% of S. aureus isolates carried the icaA gene and the detection rate of CRA was higher (36.07%) compared to the MTP test (24.59%). A total of 78.21% and 56.41% of biofilm-positive isolates were methicillin-resistant S. aureus (MRSA) and vancomycin-resistant S. aureus (VRSA), respectively. All S. aureus isolates (100%) showed multidrug resistance. The molecular analysis showed an evolutionary link between isolates and revealed a strong codon bias, three different recombination events, and positive selection in some residues of the semi-conserved segments of the icaA gene. Conclusion: The study concluded that biofilm-positive isolates have a high tendency to exhibit methicillin, vancomycin, and multidrug resistance. The findings suggest that mutation and selection are the most likely causes of codon bias in the icaA gene sequences. The variations led by recombination events and positive selection are suggestive of bacterial strategy to combat antimicrobial effects and to escape the host's immune surveillance.

Fabrication of La3+ doped Ba1-x La x TiO3 ceramics with improved dielectric and ferroelectric properties using a composite-hydroxide-mediated method.

Lanthanum (La3+) doped Ba1-x La x TiO3 (x = 0.0, 0.0025, 0.005, 0.0075) ceramics were synthesized by the composite-hydroxide-mediated method. Rietveld refinement of the XRD patterns confirmed the formation of a perovskite crystal structure that transforms from tetragonal to pseudo-cubic with La3+ doping content (x). Scanning electron microscopy displayed a dense and homogeneous microstructure with reduced grain size on La3+ doping. The frequency and temperature-dependent dielectric measurements showed an improvement in the dielectric permittivity, a decrease in the ferroelectric-paraelectric transition temperature, and an increase in the dielectric diffusivity with increasing La3+ doping content. Complex impedance analysis indicated the semiconducting behavior with a positive temperature coefficient of resistance effect, which could be explained in terms of a charge compensation mechanism in the donor doped BaTiO3. The ferroelectric hysteresis loops revealed that these ceramics are ferroelectric in nature, while an improvement in the energy storage density and energy storage efficiency was observed for the doped samples due to reduced grain size on La3+ doping. Here, the sample with x = 0.005 has a high dielectric permittivity, a low dielectric tangent loss, and the highest energy storage efficiency. This makes this composition interesting for energy storage applications.

Antimicrobial activity of essential oils of Curcuma longa and Syzygium aromaticum against multiple drug-resistant pathogenic bacteria.

The present study was conducted to investigate the antimicrobial potential of essential oils of Curcuma longa and Syzygium aromaticum against multidrug-resistant pathogenic bacteria. Four identified bacterial isolates including Methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli, Klebsiella pneumoniae, and Acinetobacter baumannii were selected and their antibiotic sensitivity was checked by disc diffusion assay. C. longa and S. aromaticum were subjected to steam distillation to obtain their essential oils. The crude essential oils were fractioned by employing column chromatography. Crude essential oils and their fractions were evaluated for their antibacterial activity by agar well diffusion assay and minimum inhibitory concentrations were calculated. All the selected bacterial isolates showed resistance to three or more than three antibiotic groups and were declared as multidrugresistant (MDRs). Crude essential oils of C. longa and S. aromaticum exhibited antimicrobial activity against all selected isolates but S. aromaticum activity was better than the C. longa with a maximum 19.3±1.50 mm zone of inhibition against A. baumannii at 1.04 µL/mL MIC. GC/MS analysis revealed the abundance of components including eugenol, eugenyl acetate, b- caryophyllene, and a- Humulene in both crude oil and fractions of S. aromaticum. While the main components of C. longa essential oil were Ar-tumerone, a-tumerone, b- Tumerone, I-Phellandrene, a-zingibirene, b- sesquiphellandrene, and p- Cymene. This study highlights that plant-based essential oils could be a promising alternative to antibiotics for which pathogens have developed resistance. C. longa and S. aromaticum carry compounds that have antimicrobial potential against multiple drug-resistant bacteria including MRSA. E. coli, K. pneumoniae and A. baumannii.

Explaining the decrease in mortality from abdominal aortic aneurysm rupture.

BACKGROUND: A steady rise in mortality from abdominal aortic aneurysm (AAA) was reported in the 1980s and 1990 s, although this is now declining rapidly. Reasons for the recent decline in mortality from AAA rupture are investigated here. METHODS: Routine statistics for mortality, hospital admissions and procedures in England and Wales were investigated. All data were age-standardized. Trends in smoking, hypertension and treatment for hypercholesterolaemia (statins), together with regression coefficients for mortality, were available from public sources for those aged at least 65 years. Deaths from ruptured AAA avoided in this age group were estimated by using the IMPACT equation: deaths avoided = (deaths in index year) × (risk factor decline) × ß-coefficient. RESULTS: From 1997, deaths from ruptured AAA have decreased sharply, almost twofold in men. Hospital admissions for elective AAA repair have increased modestly (from 40 to 45 per 100,000 population), attributable entirely to more procedures in those aged 75 years and over (P < 0.001). Admissions for ruptured AAA have declined from 18.6 to 13.5 per 100,000 population, across all ages, with the proportion offered and surviving emergency repair unchanged. From 1997, mortality from ruptured aneurysm in those aged at least 65 years has fallen from 65.9 to 44.6 per 100,000 population. An estimated 8-11 deaths per 100,000 population were avoided by a reduced prevalence of smoking and a similar number from an increase in the number of elective AAA repairs. Estimates for the effects of blood pressure and lipid control are uncertain. CONCLUSION: The reduction in incidence of ruptured AAA since 1997 is attributable largely to changes in smoking prevalence and increases in elective AAA repair in those aged 75 years and over.

Is the incidence of abdominal aortic aneurysm declining in the 21st century? Mortality and hospital admissions for England & Wales and Scotland.

BACKGROUND: Between 1951 and 1995 there was a steady increase in age-standardised deaths from all aortic aneurysms in men, from 2 to 56 per 100,000 population in England & Wales, supporting an increase in incidence. More recently, evidence from Sweden and elsewhere suggests that now the incidence of abdominal aortic aneurysm (AAA) may be declining. METHODS: National statistics for hospital admissions and deaths from AAA, after population age-standardisation, were used to investigate current trends in England & Wales and Scotland. RESULTS: Between 1997 and 2009 there has been a reduction in age-adjusted mortality from AAA from 40.4 to 25.7 per 100,000 population for England & Wales and from 30.1 to 20.8 per 100,000 population in Scotland. The decrease in mortality was more marked for men than women. Mortality decreased more than 2-fold in those <75 years versus 25% only in those >75 years. During this same time period the elective hospital admissions for AAA repair have only increased in the population >75 years. CONCLUSIONS: These data suggest that the age at which clinically-relevant aneurysms present has increased by 5-10 years and that incidence of clinically-relevant AAA in men in England & Wales and Scotland is declining rapidly. The reasons for this are unclear.

Bio-efficacy of iron and zinc fortified wheat flour along with bio-assessment of its hepatic and renal toxic potential.

Study was planned to assess the bio-efficiency along with toxicity of iron and zinc fortified whole wheat flour in Sprague dawley albino rats. Whole wheat flour was fortified with different dosage of sodium iron EDTA (NaFeEDTA), ferrous sulphate (FeSO4), zinc oxide (ZnO) and zinc sulphate (ZnSO4). The rats (n=3) in each group were fed on fortified wheat flour for 2 months. Liver biomarkers including alkaline phosphatase (ALP), alanine transaminase (ALT), aspartate aminotransferase (AST) and bilirubin were recorded from serum samples. Increased concentration of ZnSO4 affected the liver biomarkers to be highest among all whereas, bilirubin levels were less than the rats fed on control diet. The above mentioned fortificants have negligible effect on renal biomarkers including creatinine and urea. Moreover, hematological parameters were also checked and reportedly, sodium iron EDTA fed rats presented highest amount of hemoglobin, iron and total iron binding capacity. Highest zinc level was observed in rats fed on whole wheat flour fortified with 60mg/Kg Zinc oxide. Microscopic observation of liver tissue depicted that rats fed on iron and zinc fortified wheat flour have more toxic effects whereas, histopathology presentation of kidney tissue has least toxic impact. It has been concluded that mandatory fortification of wheat flour with iron and zinc may cause increased serum biomarkers along with toxicity of vital organs like liver, hence fraction of wheat flour may be fortified to fulfill the requirements of deprived and vulnerable group.

A perspective of the prevalent H9N2 virus with a special focus on molecular and pathological aspects in commercial broiler chicken in Punjab, Pakistan.

Frequent outbreaks of avian influenza H9N2 virus in Pakistan revealed that this subtype has become endemic in the poultry industry and, besides economic losses, poses a threat to public health. The present study describes the molecular characterization and pathological alterations in naturally infected broiler chickens with the current H9N2 field strain and their phylogenomic dynamics. In this study, tissue samples (trachea, lung, kidney and intestine) from 100 commercial chicken flocks were collected from July 2018 to August 2019. Samples were subjected to molecular detection, phylogeny and subsequent pathological examination. The complete length of the HA gene was successfully amplified in five samples. Nucleotide sequencing revealed positive samples placed in a clade belonging to the B2 sub-lineage of the G1 genotype and categorized as LPAIV based on the amino acid sequence of the HA gene at the cleavage site (PAKSSR/G). Genetic analysis of the haemagglutinin (HA) gene revealed nt: 80.5%-99.5%; aa: 83.8%-98.9% homology to H9N2 strains reported previously from Pakistan, neighbouring countries, and (A/Quail/Hong Kong/G1/97). Gross lesions include a slight airsacculitis, mild hemorrhages, diffuse congestion and purulent exudate in tracheal mucosa, fibrinonecrotic cast in the trachea lumen and mild pulmonary congestion. Histopathological alterations include sloughing of epithelial cells and infiltration of inflammatory cells in the trachea, mononuclear cells (MNCs) infiltration, pulmonary congestion and exudate in the lumen of parabronchi, peritubular congestion in the kidneys with degeneration of tubular epithelial cells and degenerative changes in the intestinal villi epithelial cells and goblet cell hyperplasia. Immunohistochemistry analysis confirmed the presence of AIVH9N2 antigen in the trachea, lungs, kidney and intestine. Electron microscopy revealed ultrastructural changes in the trachea, including degenerated cilia, mitochondrial swelling and enlarged endoplasmic reticulum. Based on all essential analysis, the present study revealed the distribution of the H9N2 virus of G1 genotype in Punjab, Pakistan, with mild to moderate pathogenicity.

Acaricide resistance in Boophilus microplus ticks collected from two ecological Zones of Khyber Pakhtunkhwa, Pakistan.

Boophilus microplus is a major cattle tick specie causing great economic loss to the dairy industry throughout the globe including Pakistan. Trichlorfon and Deltamethrin are used to control bovine ticks, and their sprays are also used in other pest control programs that exert pressure on ticks to gain resistance. This study is aimed to examine the resistance level of Rhipiciphalus microplus against trichlorfon and deltamethrin. The engorged ticks were collected from two ecological regions of Khyber Pakhtunkhwa, KPK Pakistan i.e., Swat & Dir (zone-1), and Charsadda & Nowshera (zone-2). Four concentrations of acaricides in two-fold and ten-fold ppm with three replicates for each were used in both bioassays. Egg hatch assay and adult immersion tests were used to assess the resistance status. The probit analysis of egg hatch assay showed the highest hatching percentage in zone 1 on both dilutions (67-76% on two-fold and 68-88% on ten-fold dilution) while lethal concentration (LC95) was found to be 2.187 ppm and discriminating dose (DD) as 4.374 ppm for trichlorfon. In zone 2, hatching percentage was 73-84 on two-fold and 72-91% on ten-fold dilution while LC95 was recorded as 0.599 ppm and DD as 1.198 ppm. The same parameters were studied for deltamethrin and in zone 1 the hatching percentage was found as 38-56% on two-fold dilution and 37-80% on ten-fold dilution while LC95 was recorded as 0.001 ppm and DD as 0.002 ppm. In zone 2, the hatchability was recorded as 42-58% on two-fold and 43-85% on ten-fold dilution. The values for LC95 was recorded as 0.001 ppm and DD as 0.002 ppm. Further, analysis of adult immersion test against trichlorfon revealed the values of LC50 as 2.85 ppm and LC95 as 4.71 ppm in zone 1 and in zone 2 as 3.14 ppm and 5.28 ppm, respectively. Similarly, LC50 and LC95 against deltamethrin was recorded as 0.79 ppm & 1.71 ppm in zone 1 and 0.45 ppm & 4.325 ppm in zone 2, respectively. Based on the findings of this study, the isolated Rhipicephalus microplus was found to be more resistant to the widely used acaricides i.e., trichlorfon than deltamethrin. In order to maintain the efficacy of acaricides at country level, the study recommends continuous monitoring of resistance.

Distribution, screening and biochemical characterization of indigenous microalgae for bio-mass and bio-energy production potential from three districts of Pakistan.

Trend of biofuel production from microalgal triacylglycerols is enhancing, because this substrate is a good sustainable and advantageous alternative to oil and gas fuel. In the present study, indigenous micro algal isolates were screened from water (n=30) and soil (n=30) samples collected from three districts of Punjab, Pakistan to evaluate their biofuel production potential. The samples were inoculated on BG - 11 agar medium plates by incubating at room temperature of 25°C providing 1000 lux for 16h light cycle followed by 8h of dark cycle for 15 d. Water samples were found to be rich in microalgae and 65.33% microalgae (49 isolates) were isolated from Faisalabad district. On the basis of microscopic morphology microalgal isolates (n=180) were selected and subjected to lipid detection by Nile red staining assay. Nile red positive isolates (n=23) were processed for biochemical (lipid, protein and carbohydrates) characterization. AIN63 isolate showed higher lipids (17.4%) content as detected by micro vanillin assay. Algal isolate AIN128 showed best protein contents (42.91%) detected by Bradford assay and AIN172 isolate showed higher carbohydrate contents (73.83%) as detected by anthrone assay. The selected algal isolates were also analyzed by Fourier transform infrared (FTIR) spectroscopy for confirmation of carbohydrate, protein and lipid analysis. These indigenous algae have the potential for in-vitro biofuel production from agricultural waste.

Antibacterial activity of plant essential oils against indigenously characterized methicillin-resistant Staphylococcus aureus (MRSA).

Plant essential oils were evaluated for antimicrobial activity against Methicillin-resistant Staphylococcus aureus (MRSA). The isolates (n=03) were procured from Institute of Microbiology, UVAS Lahore, Pakistan. After biochemical and 16S rRNA gene-based PCR characterization, accession numbers were retrieved from NCBI i.e. MW344063.1, MW344064.1 and MW344065.1. These isolates exhibited molecular positivity by multiplex PCR for mecA, coa and eta toxin genes. Moreover, these isolates exhibited resistance to cefoxitin, ampicillin, amoxicillin, penicillin, amoxicillin clavulanate, ciprofloxacin, erythromycin and gentamicin. The antibiotic resistant isolates were evaluated for antimicrobial activity of plant essential oils. The highest zone of inhibition (mean ZOI±S.D.) was measured for Cinnamomum verum (22.67±1.52 mm) followed by Eucalyptus globulus (18.67±2.51 mm) and Syzygium aromaticum (12.67±2.51 mm). Lowest mean MIC value (0.33±0.11 mg/mL) was recorded for E. globulus . Eucalyptus globulus was processed for fractionation by column chromatography and n-hexane, chloroform, n-hexane + chloroform and ethyl-acetate fractions were evaluated for antibacterial activity. Lowest mean MIC (10.04±5.80 mg/mL) was recorded for E. globulus n-hexane fraction. Cell survival percentage of BHK21 cell line was 51.7% at 54.87mg/mL concentration of E. globulus n-hexane fraction. Through gas chromatography-mass spectrometry (GC-MS) analysis of n-hexane fraction, benzene was found abundant (29.9%) as active compound. It was concluded that E. globulus n-hexane fraction exhibited significantly promising results against MRSA .

Resurgence of diphtheria in Northern Gujarat: A retrospective study done in Banas Medical College and Research Centre, Palanpur, Gujarat.

Background: A diphtheria outbreak occurred in Banaskantha district in the northern part of Gujarat in 2019-2020. This study was undertaken to study and generate information on the resurgence of the disease in this region and to generate information regarding the status of vaccination in this region and what strategies can be adopted to avoid re-emergence of the disease in the future. Materials and Methods: This is a hospital-based, retrospective, descriptive study done on patients of diphtheria who were admitted in Banas Medical College and Research Institute, Palanpur from September 2019 to January 2020. Throat swab was taken from all the patients and information regarding symptoms, vaccination, and demographic characteristic was collected. Treatment in the form of ADS, injection crystalline penicillin/erythromycin and other supportive treatment was given. Result: Out of 188 patients, 27 patients (14.36%) were less than 5 years of age, 118 patients (62.76%) and 38 patients (20.21%) were of age group 5-10 and 11-18 years, respectively. Total of five patients (2.66%) were of age more than 18 years. Out of 188 patients, 102 (54.25%) were males and 86 patients (45.74%) were females. All the patients (188) were found to be unvaccinated. Among 188 throat swabs that were tested, 21 samples (11.17%) were found to be culture positive for Corynebacterium diphtheriae. Antidiphtheric serum was given in 181 patients (96.27%) according to the requirement. Out of 188 patients, 155 patients (82.44%) improved with treatment and were discharged. Total of 23 patients (12.23%) were referred to higher center for tracheostomy and for management of other complications. Six patients (3.19%) left against medical advice and four patients (2.12%) expired despite all measures taken. Conclusion: Diphtheria is a disease that can easily be prevented by vaccination. Our study highlights need to increase awareness about vaccination among people of Banaskatha district and all measures should be taken to give full vaccination to children less than 5 years of age and booster vaccination should be encouraged in adolescents and adults so that future resurgence of disease can be prevented.