Activity of plant essential oils against antibiotic resistant Enterococcus faecalis isolated from diarrheic children.

Activity of plant essential oils and their fractions was evaluated against characterized isolates of antibiotic resistant Enterococcus faecalis recovered from diarrheic children. The isolates were confirmed by polymerase chain reaction (PCR) targeting 16S rRNA gene amplification followed by nucleotide sequencing and accession numbers retrieved were MW349990.1, MW349859.1, MW332122.1, MW356805.1, MW349975.1, MW349988.1, MW356790.1, MW356244.1, MW341593.1 and MW332549.1. These isolates were screened for antibiotic susceptibility to a wide range of antibiotic groups and mean zone of inhibition (ZOI) of all antibiotics were recorded. Antibacterial activity of plant essential oils (n=05) was checked against three antibiotic resistant isolates of E. faecalis. Three plant essential oils having higher ZOI including Cinnamomum verum, Syzygium aromaticum and Nigella sativa were used against resistant E. faecalis isolates to determine minimum inhibitory concentration (MIC). The lowest MIC observed was of S. aromaticum (11.39±3.94 mg mL-1). The S. aromaticum n-hexane plus chloroform fraction displayed higher mean ZOI (16.67±2.51 mm), while the lowest MIC was of n-hexane oil fraction. Based upon gas chromatography-mass spectrometry (GC/MS) analysis, the most effective fatty acid was eugenic acid which is present in higher proportion in both fractions. These fractions of essential oils proved safe for the treatment of antibiotic resistant diarrheic cases of children caused by E. faecalis.

An in vitro antiviral activity of iodine complexes against SARS-CoV-2.

Since the emergence of COVID-19 pandemic in China in late 2019, scientists are striving hard to explore non-toxic, viable anti-SARS-CoV-2 compounds or medicines. We determined In vitro anti-SARS-CoV-2 activity of oral formulations (syrup and capsule)of an Iodine-complex (Renessans). First, cell cytotoxicity of Renessans on the Vero cells was determined using MTT assay. Afterwards, the antiviral activity of Renessans was determined using viral inhibition assays and TCID50. For this, nontoxic concentrations of the Renessans were used. The results showed that Renessans is nontoxic to the cells up to 50 µg/mL. At 1.5 µg/mL concentration, SARS-CoV-2 production was significantly reduced to 101.43 TCID50 and 101.58 TCID50 for the syrup and capsule, respectively, as compare to virus infected control cells 106.08 TCID50 and we found the dose dependent inhibition of virus replication in the presence of Renessans. Renessans inhibited SARS-CoV-2 with an EC50 value of 0.425 µg/mL and 0.505 µg/mL for syrup and capsule, respectively. Furthermore, there was no virus detected at concentration of 50 µg/mL of Renessans. This study indicates that Renessans, containing iodine, have potential activity against SARS-CoV-2 which needs to be further investigated in human clinical trials.

Appraisal of anti-mycobacterial potential against MDR-MTB in pediatric patients, cytotoxicity and mutagenicity of Aloe vera and Allium sativum.

The purpose of the studies was to evaluate an in-vitro anti-mycobacterial activity of Aloe vera and Allium sativum against MDR-MTB, their cytotoxicity and mutagenicity. Four extracts of Aloe vera and Allium sativum were prepared by Soxhlet apparatus and their minimum inhibitory concentrations (MIC's) were determined by BACTEC MGIT960 system against multi drug resistant Mycobacterium tuberculosis (MDR-MTB) isolates, collected from pediatric patients. Fractions of Aloe vera and Allium sativum extracts were separated using glass column chromatography, followed by evaluation of cytotoxicity and mutagenicity by tetrazolium salt (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and Ames test, respectively. Out of four extracts, ethanol extracts of Aloe vera and Allium sativum exhibited activity at MIC 5mg/mL to 7mg/mL and 3mg/mL to 5mg/mL, respectively and IC50 by MTT assay for combination of all fractions were 278.3mcg/100µL and 270.8mcg/100µL and in Ames assay M.I of TA98 were 0.14 and 0.07 and M.I of TA100 were 1.14 and 0.44, respectively. Aloe vera and Allium sativum extracts showed anti-mycobacterial activity against MDR-MTB isolates so, MIC of ethanol extracts of each plant and fractions of column chromatography had been checked. The MTT and Ames tests depicted that ethanol extracts of Aloe vera and Allium sativum were non-cytotoxic and non-mutagenic, and can be used in treatment of patients suffering from MDR-MTB.

Immunoprotective role of LaSota vaccine under immunosuppressive conditions in chicken challenged with velogenic avian avulavirus-1.

The first objective of the present study was to evaluate if the antibodies induced by the live LaSota and killed Newcastle disease (sub-genotype VIIi) vaccines protect the chickens against exposure with pathogenic avian avulavirus-1 (AAvV-1) of chicken and/or pigeon origins. The second objective was to study the effect of vaccines on stressed birds (dexamethasone, aflatoxin, and heat stressed) with respect to antibody production and protection against pathogenic AAvV-1 challenge. Sixty-one-day-old Hubbard chickens were divided into six groups (gA-gF) with ten animals each. All the groups received LaSota (105 EID50, 0.1 ml per chick) on days 7 and 27 via eye drop and one intramuscular injection of a killed vaccine (sub-genotype VIIi) (107.5 EID50, 1 ml) on day 18, except the control birds received the PBS only. Moreover, group gC-DEX received dexamethasone intramuscularly at a dose rate of 1-mg/kg body weight daily; gD-AFLA had received aflatoxin as oral gavage at a dose rate of 30 ppb daily, and gE-HEAT was kept under heat stressed (38 °C) till challenged. All the groups were challenged with AAvV-1 strain of chicken origin of sub-genotype VIIi, except the group gA-pigeon was challenged with pigeon-origin strain (sub-genotype VIm). The result showed that the gA-pigeon and gB-chicken vaccinate showed 100% and 80% protection. The immunosuppressive birds produced low pre-challenge HI titer, and protection was observed at 40%, 50%, and 70% in gC-DEX, gD-AFLA, and gE-HEAT, respectively. Our findings suggest the stress factors such as aflatoxin in the feed and dexamethasone are immunosuppressive in nature and suppress the immune response and its associated protective role during infection.

Antiviral, embryo toxic and cytotoxic activities of Astragalus membranaceus root extracts.

Antiviral activity of Astragalus membranaceus aqueous and methanol root extracts was determined against Avian influenza H9 virus. Toxicity profile of extracts was evaluated using chicken embryos and BHK-21 cell line. Different concentrations (400, 200, 100, 50, 25. 12.5, 6.25 and 3.12µg/mL) of both aqueous and methanol extracts were mixed with standard virus inoculum (4HAunits) and incubated for 30minutes at 37°C prior to inject the chicken embryos. Chorioallantoic fluid harvested 72 hours post inoculation and evaluated for virus growth using hemagglutination assay. Same concentrations of both extracts without virus were injected in chicken embryos to evaluate embryo toxic activity as well. The cytotoxic activity of aqueous and methanol extracts was determined by MTT colorimetric assay using BHK-21 cells. Three concentrations (400, 200 and 100µg/mL) of aqueous and five concentrations (400, 200, 100, 50 and 25µg/mL) of methanol extract showed antiviral activity. None of the tested concentrations of aqueous and methanol A. membranaceus root extracts caused chicken embryo mortality. Cell survival percentage of aqueous extract was higher than 50 at all of the tested concentrations except 400µg/mL. Two concentrations (400 and 200µg/mL) of methanol extract showed cytotoxicity. It was concluded that aqueous and methanol roots extracts of A. membranaceus have antiviral activity and concentrations which were safe may be used for treatment of Avian influenza H9 virus infections.

Role of Rab GTPases in HSV-1 infection: Molecular understanding of viral maturation and egress.

Most enveloped viruses exploit complex cellular pathways for assembly and egress from the host cell, and the large DNA virus Herpes simplex virus 1 (HSV-1) makes no exception, hijacking several cellular transport pathways for its glycoprotein trafficking and maturation, as well as for viral morphogenesis and egress according to the envelopment, de-envelopment and re-envelopment model. Importantly Rab GTPases, widely distributed master regulators of intracellular membrane trafficking pathways, have recently being tightly implicated in such process. Indeed, siRNA-mediated genetic ablation of specific Rab proteins differently affected HSV-1 production, suggesting a complex role of different Rab proteins in HSV-1 life cycle. In this review, we discuss how different Rabs can regulate HSV-1 assembly/egress and the potential therapeutic applications of such findings for the management of HSV-1 infections.

Anti-avian influenza virus H9N2 activity of aqueous extracts of Zingiber officinalis (Ginger) and Allium sativum (Garlic) in chick embryos.

In the present study, anti-Avian influenza virus H9N2 activity of aqueous extracts (5, 10, 15, 20, 25%) of Zingiber officinalis and Allium sativum was evaluated. Embryo-toxicity was evaluated by histopathological scoring of Chorio-allantoic membrane of chick embryos. Cytotoxicity of extracts was determined by MTT assay on Vero cells. Aqueous extract of ginger had antiviral activity at 10, 15, 20 and 25% while garlic had activity at 15, 20 and 25%. Histopathological scoring of chorio-allantoic membrane for aqueous extracts (5, 10, 15, 20, 25%) of ginger (0.66±0.57, 1.33±0.57, 1.66±0.57, 2.66±0.57, 3.66±0.57, respectively) and garlic (1.00±0.00, 1.33±0.57, 2.00±0.00, 2.33±0.57, 3.66±0.57, respectively) was concentration dependant. MTT assay revealed cytotoxicity of both plants was also concentration dependent. Extracts of ginger (5, 10, 15, 20, 25%) had lower cytotoxicity (71, 59, 28, 22, 0 % cell survival, respectively) as compared to garlic (61, 36. 20, 11, 3% cell survival, respectively). Overall results revealed that concentration of aqueous extract of ginger (10%), showing antiviral activity against H9N2, was less toxic to vero cells (> 50% cell survival). It is insinuated that ginger may have anti- Avian influenza virus H9N2 potential and its active compounds needs further investigations.

Safety climate in Pakistani universities' laboratories: an assessment of individual factors.

Objectives. Current research aims to identify factors that affect the occupational safety climate in university laboratories despite their perception as low-risk areas compared to industrial environments. Methods. A safety climate survey was conducted in science laboratories across various engineering universities in Pakistan. The survey questionnaire was administered to 406 personnel, and a quantitative method for analysis was selected to examine the socio-demographic variables. A 5-point Likert scale (1 = strongly disagree to 5 = strongly agree) was used to perceive responses from participants. Additionally, a scale reliability test was conducted, and multivariate analysis of variance was performed to determine the relationship between selected dependent and independent variables. Results. The study found an overall safety climate score of 3.16 ± 0.55, indicating a moderate to high perception of safety on a scale of 1-5. Parameters such as role in the laboratory, departments/disciplines, accident experience and safety training significantly affected the safety climate score, while gender, age group, duration in university and accident witnessing did not. Conclusion. Upper management involvement, safety communication and direct supervision are crucial for improving the safety climate of university laboratories. The study recommends the consideration of the identified significant safety climate dimensions in laboratory safety policy-making at academic institutes.

Machining of Triangular Holes in D2 Steel by the Use of Non-Conventional Electrodes in Die-Sinking Electric Discharge Machining.

Electric discharge machining is relatively a slow process in terms of machining time and material removal rate. The presence of overcut and the hole taper angle caused by the excessive tool wear are other challenges in the electric discharge machining die-sinking process. The areas of focus to solve these challenges in the performance of electric discharge machines include increasing the rate of material removal, decreasing the rate of tool wear, and reducing the rate of hole taper angle and overcut. Triangular cross-sectional through-holes have been produced in D2 steel through die-sinking electric discharge machining (EDM). Conventionally, the electrode with uniform triangular cross-section throughout the electrode length is used to machine triangular holes. In this study, new designs of electrodes (non-conventional designs) are employed by introducing circular relief angles. For material removal rate (MRR), tool wear rate (TWR), overcut, taper angle, and surface roughness of the machined holes, the machining performance of conventional and unconventional electrode designs is compared. A significant improvement in MRR (32.6% increase) has been achieved by using non-conventional electrode designs. Similarly, the hole quality resulted by non-conventional electrodes is way better than hole quality corresponding to conventional electrode designs, especially in terms of overcut and hole taper angle. A reduction of 20.6% in overcut and a reduction of 72.5% in taper angle can be achieved through newly designed electrodes. Finally, one electrode design has been selected (electrode with 20 degree relief angle) as the most appropriate electrode resulting in better EDM performance in terms of MRR, TWR, overcut, taper angle, and surface roughness of triangular holes.

Electric Discharge Machining of Ti6Al4V ELI in Biomedical Industry: Parametric Analysis of Surface Functionalization and Tribological Characterization.

The superior engineering properties and excellent biocompatibility of titanium alloy (Ti6Al4V) stimulate applications in biomedical industries. Electric discharge machining, a widely used process in advanced applications, is an attractive option that simultaneously offers machining and surface modification. In this study, a comprehensive list of roughening levels of process variables such as pulse current, pulse ON time, pulse OFF time, and polarity, along with four tool electrodes of graphite, copper, brass, and aluminum are evaluated (against two experimentation phases) using a SiC powder-mixed dielectric. The process is modeled using the adaptive neural fuzzy inference system (ANFIS) to produce surfaces with relatively low roughness. A thorough parametric, microscopical, and tribological analysis campaign is established to explore the physical science of the process. For the case of the surface generated through aluminum, a minimum friction force of ~25 N is observed compared with the other surfaces. The analysis of variance shows that the electrode material (32.65%) is found to be significant for the material removal rate, and the pulse ON time (32.15%) is found to be significant for arithmetic roughness. The increase in pulse current to 14 A shows that the roughness increased to ~4.6 µm with a 33% rise using the aluminum electrode. The increase in pulse ON time from 50 µs to 125 µs using the graphite tool resulted in a rise in roughness from ~4.5 µm to ~5.3 µm, showing a 17% rise.

Temporal study of wastewater surveillance from September 2020 to March 2021: an estimation of COVID-19 patients in Lahore, Pakistan.

The first aim of study was to quantify the viral load in the wastewater samples by RT-qPCR testing in Lahore population to estimate the number of patients affected and predict the next resurgence of COVID-19 wave in the city. The second aim of the study was to determine the hotspot areas of Lahore which remained positive more often for virus with high viral load. In this study, n = 420 sewage samples were collected on an average of two weeks intervals from 30 different sewage water disposal stations (14 sampling events) from Sept 2020 to March 2021. RNA was extracted and quantified by RT-qPCR without concentrating the virus in samples. Number of positive disposal sites (7-93%), viral load from sewage samples (100.296 to 103.034), and estimated patients (660-17,030) ranged from low to high according to the surge and restrain of 2nd and 3rd COVID-19 waves in the country. The viral load and estimated patients were reported high in January 2021 and March 2021 which were similar to the peak of 2nd and 3rd waves in Pakistan. Site 18 (Niaz Baig village DS) showed the highest viral load among all sites. Findings of the present study helped to estimate the number of patients and track the resurgence in COVID-19 waves in Lahore particularly, and in Punjab generally. Furthermore, it emphasizes the role of wastewater-based epidemiology to help policymakers strengthen the quarantine measures along with immunization to overcome enteric viral diseases. Local and national stake holders should work in collaboration to improve the environmental hygiene to control the disease.

Molecular Characterization and Selection of Indigenous SARS-CoV-2 Delta Variant for the Development of the First Inactivated SARS-CoV-2 Vaccine of Pakistan.

Vaccines are one of the efficient means available so far for preventing and controlling the infection rate of COVID-19. Several researchers have focused on the whole virus's (SARS-CoV-2) inactivated vaccines which are economically efficient to produce. In Pakistan, multiple variants of SARS-CoV-2 have been reported since the start of the pandemic in February 2020. Due to the continuous evolution of the virus and economic recessions, the present study was designed to develop an indigenous inactivated SARS-CoV-2 vaccine that might help not only to prevent the COVID-19 in Pakistan, it will also save the country's economic resources. The SARS-CoV-2 were isolated and characterized using the Vero-E6 cell culture system. The seed selection was carried out using cross-neutralization assay and phylogenetic analysis. The selected isolate of SARS-CoV-2 (hCoV-19/Pakistan/UHSPK3-UVAS268/2021) was inactivated using beta-propiolactone followed by vaccine formulation using Alum adjuvant, keeping the S protein concentration as 5 µg/dose. The vaccine efficacy was evaluated by in vivo immunogenicity testing in laboratory animals and in in vitro microneutralization test. The phylogenetic analysis revealed that all the SARS-CoV-2 isolates reported from Pakistan nested into different clades, representing multiple introductions of the virus into Pakistan. The antisera raised against various isolates from different waves in Pakistan showed a varied level of neutralization titers. However, the antisera produced against a variant (hCoV-19/Pakistan/UHSPK3-UVAS268/2021; fourth wave) efficiently neutralized (1:64-1:512) all the tested SARS-CoV-2 isolates. The inactivated whole virus vaccine of SARS-CoV-2 was safe and it also elicited a protective immune response in rabbits and rhesus macaques on the 35th-day post-vaccination. The activity of neutralizing antibodies of vaccinated animals was found at 1:256-1:1024 at 35 days post-vaccination, indicating the effectiveness of the double-dose regime of the indigenous SARS-CoV-2 vaccine.

A Novel Flushing Mechanism to Minimize Roughness and Dimensional Errors during Wire Electric Discharge Machining of Complex Profiles on Inconel 718.

One of the sustainability goals in the aeronautical industry includes developing cost-effective, high-performance engine components possessing complex curved geometries with excellent dimensional precision and surface quality. In this regard, several developments in wire electric discharge machining have been reported, but the influence of flushing attributes is not thoroughly investigated and is thus studied herein. The influence of four process variables, namely servo voltage, flushing pressure, nozzle diameter, and nozzle-workpiece distance, were analyzed on Inconel 718 in relation to geometrical errors (angular and radial deviations), spark gap formation, and arithmetic roughness. In this regard, thorough statistical and microscopical analyses are employed with mono- and multi-objective process optimization. The grey relational analysis affirms the reduction in the process's limitations, validated through confirmatory experimentation results as 0.109 mm spark gap, 0.956% angular deviation, 3.49% radial deviation, and 2.2 µm surface roughness. The novel flushing mechanism improved the spark gap by 1.92%, reducing angular and radial deviations by 8.24% and 29.11%, respectively.

Renessans Helps in Early Clearance of SARS-CoV-2: In-Vivo Activity of the Iodine Complex in Rhesus macaque.

Iodine complexes have known antimicrobial properties along with reported in-vitro antiviral activity for several viruses. Renessans is one such product with iodine complexes and ascorbic acid. The present study was designed to determine its efficacy for SARS-CoV-2 in Rhesus macaque. Rhesus macaque were assigned to: A) prophylactic group (n = 3), (B) treatment group (n = 3), (C) infection control group (n = 4), and (D) negative control group (n = 4). Groups A, B, and C were challenged with 2 × 106 TCID of SARS-CoV-2. The prophylactic group (A) was administered Renessans from 5 days before infection till 8 days postinfection (DPI). The treatment group (B) was administered Renessans from 3 till 8 DPI. Group C was administered water-insoluble fractions only. Nasal swabs from all monkeys of groups A, B, and C remained positive for SARS-CoV-2 till 2 and 7 DPI, while the swabs became negative for groups A and B at 14 DPI. Likewise, fecal matter of monkeys in group A returned negative results during the experiment, while that of group B had significantly decreased viral load (101.5 genome copies/mL) compared to group C (103 genome copies/mL). Hence, it is concluded that Renessans has in-vivo SARS-CoV-2 activity and may result in early clearance of SARS-CoV-2.

A Cross-Sectional Survey of Knowledge, Attitude, and Practices of University Students in Pakistan Regarding COVID-19.

The COVID-19 pandemic is striking the world with serious public health and socioeconomic complications. The pandemic has influenced all forms of daily life, including educational institutions. Therefore, this cross-sectional survey was conducted to understand knowledge, attitudes, and practices related to COVID-19 among the students of the University of Veterinary and Animal Sciences, Lahore. The data was collected using an online self-directed questionnaire. The survey form includes six items about sociodemographic characteristics, 14 knowledge-based questions, seven questions on attitude, and eight questions on practices. The sample number was calculated using the Raosoft sample size calculator. A total number of 3,854 students, including 1,823 men and 2,031 women, were engaged in this survey, having student representation from all the provinces in the country. The data were analyzed using a chi-square test. A total of 97% of the students knew that the etiological agent of COVID-19 is a virus and that it is a disease of the respiratory system (94%). Many students kept visiting their relatives during the lockdown (45%), and their relatives kept visiting them at home (59%). The responses from the students varied a lot on specific questions about the transmission of the virus. Women tended to have less information regarding precautionary travel measures (p < 0.01), but supplemental knowledge of prevention of disease transmission from positive patients (p < 0.01). Conclusively, the majority of the university students surveyed had imperative knowledge, a good attitude, and active practice in response to the COVID-19 outbreak. Moreover, the KAP scores have varied by demography, gender, and the number of family members. Therefore, continuous awareness of preventative behaviors should be disseminated regularly in emergencies.

Modelling the Mechanical Attributes (Roughness, Strength, and Hardness) of Al-alloy A356 during Sand Casting.

Sand-casting is a well established primary process for manufacturing various parts of A356 alloy. However, the quality of the casting is adversely affected by the change in the magnitude of the control variables. For instance, a larger magnitude of pouring velocity induces a drop effect and a lower velocity increases the likelihood of cold-shut and mis-run types of defects. Similarly, a high pouring temperature causes the formation of hot tears, whereas a low temperature is a source of premature solidification. Likewise, a higher moisture content yields microcracks (due to gas shrinkages) in the casting and a lower moisture content results in the poor strength of the mold. Therefore, the appropriate selection of control variables is essential to ensure quality manufactured products. The empirical relations could provide valuable guidance in this regard. Additionally, although the casting process was optimized for A356 alloy, it was mostly done for a single response. Therefore, this paper aimed to formulate empirical relations for the contradictory responses, i.e., hardness, ultimate tensile strength and surface roughness, using the response surface methodology. The experimental results were comprehensively analyzed using statistical and scanning electron microscopic analyses. Optimized parameters were proposed and validated to achieve castings with high hardness (84.5 HB) and strength (153.5 MPa) with minimum roughness (5.8 µm).

Evaluating Material's Interaction in Wire Electrical Discharge Machining of Stainless Steel (304) for Simultaneous Optimization of Conflicting Responses.

Stainless steel (SS 304) is commonly employed in industrial applications due to its considerable corrosion resistance, thermal resistance, and ductility. Most of its intended applications require the formation of complex profiles, which justify the use of wire electrical discharge machining (WEDM). However, its high thermal resistance imposes a limitation on acquiring adequate surface topography because of the high surface tension of the melt pool, which leads to the formation of spherical modules; ultimately, this compromises the surface quality. Furthermore, the stochastic nature of the process makes it difficult to optimize its performance, especially if more than one conflicting response is involved, such as high cutting speed with low surface roughness and kerf width. Therefore, this study aimed to comprehensively investigate the interaction of SS 304 and WEDM, with a prior focus on simultaneously optimizing all the conflicting responses using the Taguchi-based grey relational approach. Analysis of variance (ANOVA) revealed that the current was the most significant parameter for cutting speed and kerf, whereas roughness, voltage (45%), drum speed (25.8%), and nozzle offset distance (~21%) were major contributing factors. SEM micrographs showed that optimal settings not only ensured simultaneous optimization of the conflicting responses but also reduced the number and size of spherical modules.

Genetic Diversity of Canine Babesia Species Prevalent in Pet Dogs of Punjab, Pakistan.

Canine babesiosis is a serious threat to dogs' health worldwide, caused by the intra-erythrocytic Babesia species. The present study was carried out in pet dogs presented at three clinics of Lahore and one of Narowal in Punjab, Pakistan. Two hundred blood samples (50 from each clinic) were collected and screened by microscopy for Babesia spp. Out of 200 samples, 84 (42%) were found to be positive for babesiosis. The highest number of positive cases (50%) was recorded in dogs at Narowal clinic. Non-significant variation (p > 0.05) was observed in the prevalence of babesiosis in dogs in relation to sex and age. Positive samples were further confirmed by a polymerase chain reaction using 18S-rRNA genus-specific and species-specific primers. Amplicons were further analyzed by nucleotide sequencing for genetic diversity. Babesia canis and gibsoni were confirmed by genome sequencing in all diseased dogs. These isolates closely resembled each other, but differed from previous reported strains. In conclusion, pet dogs suffering from babesiosis were infected with B. canis and gibsoni, while in other countries, other Babesia species are also prevalent.

Cross Sectional Study and Risk Factors Analysis of Francisella tularensis in Soil Samples in Punjab Province of Pakistan.

Tularemia is an endemic zoonotic disease in many parts of the world including Asia. A cross-sectional study was conducted to determine genome-based prevalence of Francisella tularensis (Ft) in soil, assess an association between its occurrence in soil and likely predictors i.e., macro and micro-nutrients and several categorical variables, and determine seroconversion in small and large ruminants. The study included a total of 2,280 soil samples representing 456 villages in eight districts of the Punjab Province of Pakistan followed by an analysis of serum antibodies in 707 ruminants. The genome of Ft was detected in 3.25% (n = 74, 95% CI: 2.60-4.06) of soil samples. Soluble salts (OR: 1.276, 95% CI: 1.043-1.562, p = 0.015), Ni (OR: 2.910, 95%CI: 0.795-10.644, p = 0.106), Mn (OR:0.733, 95% CI:0.565-0.951, p = 0.019), Zn (OR: 4.922, 95% CI:0.929-26.064, p = 0.061) and nutrients clustered together as PC-1 (OR: 4.76, 95% CI: 2.37-9.54, p = 0.000) and PC-3 (OR: 0.357, 95% CI: 0.640, p = 0.001) were found to have a positive association for the presence of Ft in soil. The odds of occurrence of Ft DNA in soil were higher at locations close to a water source, including canals, streams or drains, [χ2 = 6.7, OR = 1.19, 95% CI:1.05-3.09, p = 0.004] as well as places where animals were present [χ2 = 4.09, OR = 2.06, 95% CI: 1.05-4.05, p = 0.02]. The seroconversion was detected in 6.22% (n = 44, 95% CI: 4.67-8.25) of domestic animals. An occurrence of Ft over a wide geographical region indicates its expansion to enzootic range, and demonstrates the need for further investigation among potential disease reservoirs and at-risk populations, such as farmers and veterinarians.

Seroprevalence and risk factors of glanders in working equines - Findings of a cross-sectional study in Punjab province of Pakistan.

Glanders is an infectious and contagious bacterial disease of equines. A little is known about its seroprevalence and risk factors in working equines in countries where the disease is endemic. Also, there are no reports on prevalence of the disease in areas where there is a prior evidence of Burkholderia (B.) mallei detection in soil. A cross-sectional study was conducted in selected districts (n=09) of Punjab province of Pakistan during 2014-2015. A total of 1008 serum samples were screened for detection of antibodies to B. mallei with complement fixation test followed by western blot. The overall seroprevalence was found to be 3.17% (95% CI: 2.25-4.44). The seropositivity was significantly higher from the sampling sites where B. mallei was detected in soil [OR: 10.66 (95% CI: 4.42-31.66), p=0.00]. Other risk factors significantly associated with animal seropositivity were: age group [OR: 1.78 (95% CI: 4.58-15.56), p=0.00], location in urban area [OR: 2.99 (95% CI: 1.46-6.51), p=0.00],body condition [OR: 3.47 (95% CI: 1.64-7.99), p=0.00], presence of farcy lesion[OR: 7.71 (95% CI: 3.47-19.50), p=0.00], proximity to water bodies [OR: 7.71 (95% CI: 3.47-19.50), p=0.00]; domestic animal population [OR: 3.20 (95% CI: 1.24-10.87), p=0.03] and number of households in sampling area [OR: 4.18 (95%CI: 1.82-11.30), p=0.00]. The study provides an estimate of prevalence of glanders and a potential link between animal seropositivity and presence of B. mallei in soil. The risk factors identified in this study can be used in surveillance and disease awareness. The high prevalence of disease in draught horses and contact of infected animals with their care-takers in developing countries signify need to initiate progressive control of the disease using one health approach.