Characterisation of drug-resistant Mycobacterium tuberculosis mutations and transmission in Pakistan

Tuberculosis, caused by Mycobacterium tuberculosis, is a high-burden disease in Pakistan, with multi-drug (MDR) and extensive-drug (XDR) resistance, complicating infection control. Whole genome sequencing (WGS) of M. tuberculosis is being used to infer lineages (strain-types), drug resistance mutations, and transmission patterns—all informing infection control and clinical decision making. Here we analyse WGS data on 535 M. tuberculosis isolates sourced across Pakistan between years 2003 and 2020, to understand the circulating strain-types and mutations related to 12 anti-TB drugs, as well as identify transmission clusters. Most isolates belonged to lineage 3 (n = 397; 74.2%) strain-types, and were MDR (n = 328; 61.3%) and (pre-)XDR (n = 113; 21.1%). By inferring close genomic relatedness between isolates (< 10-SNPs difference), there was evidence of M. tuberculosis transmission, with 55 clusters formed consisting of a total of 169 isolates. Three clusters consist of M. tuberculosis that are similar to isolates found outside of Pakistan. A genome-wide association analysis comparing ‘transmitted’ and ‘non-transmitted’ isolate groups, revealed the nusG gene as most significantly associated with a potential transmissible phenotype (P = 5.8 × 10–10). Overall, our study provides important insights into M. tuberculosis genetic diversity and transmission in Pakistan, including providing information on circulating drug resistance mutations for monitoring activities and clinical decision making

Assessment of Geographical Distribution of Emerging Zoonotic Toxoplasma gondii Infection in Women Patients Using Geographical Information System (GIS) in Various Regions of Khyber Pakhtunkhwa (KP) Province, Pakistan

Toxoplasmosis is a zoonotic parasitic disease caused by T. gondii, an obligate intracellular apcomplexan zoonotic parasite that is geographically worldwide in distribution. The parasite infects humans and all warm-blooded animals and is highly prevalent in various geographical regions of the world, including Pakistan. The current study addressee prevalence of Toxoplasma infection in women in various geographical regions, mapping of endemic division and t district of Khyber Pakhtunkhwa province through geographical information system (GIS) in order to locate endemic regions, monitor seasonal and annual increase in prevalence of infection in women patients. Setting: Tertiary hospitals and basic health care centers located in 7 divisions and 24 districts of Khyber Pakhtunkhwa (KP) province of Pakistan. During the current study, 3586 women patients from 7 divisions and 24 districts were clinically examined and screened for prevalence of T. gondii infection. Participants were screened for Toxoplasma infection using ICT and latex agglutination test (LAT) as initial screening assay, while iELISA (IgM, IgG) was used as confirmatory assay. Mapping of the studied region was developed by using ArcGIS 10.5. Spatial analyst tools were applied by using Kriging/Co-kriging techniques, followed by IDW (Inverse Distance Weight) techniques. Overall prevalence of T. gondii infection was found in 881 (24.56%) patients. A significant (&lt;0.05) variation was found in prevalence of infection in different divisions and districts of the province. Prevalence of infection was significantly (&lt;0.05) high 129 (30.07%) in Kohat Division, followed by 177 (29.06%), 80 (27.87%), 287 (26.72%), 81 (21.21%), 47 (21.07%), and 80 (13.71%) cases in Hazara Division, D.I Khan Division, Malakand Division, Mardan Division, Bannu Division, and Peshawar Division. Among various districts, a significant variation (&lt;0.05) was found in prevalence of infection. Prevalence of infection was significantly (&lt;0.05) high 49 (44.95%) in district Karak, while low (16 (10.81%) in district Nowshera. No significant (&gt;0.05) seasonal and annual variation was found in prevalence of Toxoplasma infection. LAT, ICT and ELISA assays were evaluated for prevalence of infection, which significantly (&lt;0.05) detected T. gondii antibodies. LAT, ICT and ELISA assays significantly (&lt;0.05) detected infection, while no significant (&gt;0.05) difference was found between positivity of LAT and ICT assays. A significant difference (&lt;0.05) was found in positivity of Toxoplasma-specific (IgM), (IgG) and (IgM, IgG) immunoglobulin by ICT and ELISA assay. The current study provides comprehensive information about geographical distribution, seasonal and annual variation of Toxoplasmosis infection in various regions of Khyber Pakhtunkhwa province of Pakistan. Infection of T. gondii in women shows an alarming situation of disease transmission from infected animals in the studied region, which is not only a serious and potential threat for adverse pregnancy outcomes, but also cause socioeconomic burden and challenges for various public and animal health organizations in Pakistan and across the country

Isolation and Identification of Common Contaminants Bacteria from Working Area in Microbiology Laboratory

Abstract: Frequent environmental contaminants within microbiology laboratory create not only diagnostic quandaries but also poses major risk for health care workers and patients. Objective of our study was to isolate and identify the common laboratory contaminant bacteria with an ultimate goal to reduce false positive culture reports as well as Laboratory acquired infections. The study was conducted in laboratory of Microbiology Department Kohat University of Science and Technology Kohat from March 2014 to June 2014. Total 5 samples were collected from different areas of laboratory including table, floor, clothing, air, and incubator. Out of 5 collected samples, growth was observed. Out of 5 culture positive samples total 22 bacterial contaminants were isolated and identified by various biochemical tests. Out of 22 bacterial contaminants, 8 (36.36%) was Staphylococcus epidermis&apos;s which was the most common contaminants. 7 (31.81%) was Bacillus subtilis which was the second most common contaminants. 4 (18.18%) was Staphylococcus aerous while 3 (13.63%) was Deptheriods which showed minimum bacterial contaminants. Precaution should be taken to get rid of these organisms from laboratory by means of proper laboratory disinfection and sterilization as well as personal hygiene of laboratory workers. From our study we conclude that laboratory can be a potential source of contamination to give false positive results

Honey: Single food stuff comprises many drugs

Honey is a natural food item produced by honey bees. Ancient civilizations considered honey as a God gifted prestigious product. Therefore, a huge literature is available regarding honey importance in almost all religions. Physically, honey is a viscous and jelly material having no specific color. Chemically, honey is a complex blend of many organic and inorganic compounds such as sugars, proteins, organic acids, pigments, minerals, and many other elements. Honey use as a therapeutic agent is as old as human civilization itself. Prior to the appearance of present day drugs, honey was conventionally used for treating many diseases. At this instant, the modern research has proven the medicinal importance of honey. It has broad spectrum anti-biotic, anti-viral and anti-fungal activities. Honey prevents and kills microbes through different mechanism such as elevated pH and enzyme activities. Till now, no synthetic compound that works as anti-bacterial, anti-viral and anti-fungal drugs has been reported in honey yet it works against bacteria, viruses and fungi while no anti-protozoal activity has been reported. Potent anti-oxidant, anti-inflammatory and anti-cancerous activities of honey have been reported. Honey is not only significant as anti-inflammatory drug that relieve inflammation but also protect liver by degenerative effects of synthetic anti-inflammatory drugs. This article reviews physico-chemical properties, traditional use of honey as medicine and mechanism of action of honey in the light of modern scientific medicinal knowledge

Resistance to Cypermethrin Is Widespread in Cattle Ticks (Rhipicephalus microplus) in the Province of Punjab, Pakistan: In Vitro Diagnosis of Acaricide Resistance

Control of the cattle tick Rhipicephalus (R.) microplus mainly relies on chemical acaricides and cypermethrin is the most widely used acaricide in Pakistan. Farmers frequently complain about its low efficacy, thus, the present study was designed to quantify the frequency of cypermethrin resistance in cattle ticks. Engorged female R. microplus were collected and tested for the efficacy of cypermethrin using the FAO-recommended larval packet test. Resistance factors (RF) were estimated at both the lethal concentration for 50% (LC50) and 99% (LC99) of ticks. Thirty-three samples were tested, of which 8/33 (24.24%) were classified as resistant based on the RF50, and all 33 were classified as resistant based on the RF99. In District Sargodha, when only the RF50 was considered, 45.5% of samples were classified as resistant, but at RF99, all tested samples were identified as resistant. In District Okara, the variation in RF50 estimates was 2.2&ndash;8.3 and variation in RF99 estimates was 10.6&ndash;1139.8. Similar results were found in District Attock, where variations in RF50 were 0.8&ndash;8.5 and RF99 ranged from 9&ndash;237.3. The study showed that cypermethrin resistance is prevalent in these three districts of Pakistan and is likely to be overestimated by classification based on the RF99

Antinematode Activity of Abomasum Bacterial Culture Filtrates against Haemonchus contortus in Small Ruminants

Haemonchosis is a parasitic disease of small ruminants that adversely affects livestock production. Haemonchus contortus is one of the most prevalent nematode parasites that infect the abomasum of small ruminants. This parasite reduces milk production, overall growth and sometimes causes the death of the infected animals. The evaluation of the biocontrol potential of some abomasum bacterial isolates against H. contortus is investigated in this study. Out of which, three isolates—Comamonas testosteroni, Comamonas jiangduensis, Pseudomonas weihenstephanesis—show significant effect against the nematode L3, adult, and egg hatch inhibition assays. Various concentrations of metabolites from these bacteria are prepared and applied in different treatments compared with control. In the case of adult mortality assay, 50% metabolites of C. testosteroni and P. weihenstephanesis show 46% adult mortality, whereas C. jiangduensis shows 40% mortality. It is observed that decreasing the concentration of bacterial metabolite, lowers nematode mortality. The minimum nematode mortality rate is recorded at the lowest filtrates concentration of all the bacterial isolates. The same trend is observed in egg hatch inhibition assay, where the higher concentration of bacterial culture filtrates shows 100% inhibition of H. contortus egg. It is concluded that the effect of bacterial culture filtrates against H. contortus is dose-dependent for their activity against nematode L3, adult, and inhibition of egg hatchment