A Study of Ticks and Tick-Borne Livestock Pathogens in Pakistan

Background As obligate blood-feeding arthropods, ticks transmit pathogens to humans and domestic animals more often than other arthropod vectors. Livestock farming plays a vital role in the rural economy of Pakistan, and tick infestation causes serious problems with it. However, research on tick species diversity and tick-borne pathogens has rarely been conducted in Pakistan. In this study, a systematic investigation of the tick species infesting livestock in different ecological regions of Pakistan was conducted to determine the microbiome and pathobiome diversity in the indigenous ticks. Methodology/Principal findings A total of 3,866 tick specimens were morphologically identified as 19 different tick species representing three important hard ticks, Rhipicephalus, Haemaphysalis and Hyalomma, and two soft ticks, Ornithodorus and Argas. The bacterial diversity across these tick species was assessed by bacterial 16S rRNA gene sequencing using a 454-sequencing platform on 10 of the different tick species infesting livestock. The notable genera detected include Ralstonia, Clostridium, Staphylococcus, Rickettsia, Lactococcus, Lactobacillus, Corynebacterium, Enterobacter, and Enterococcus. A survey of Spotted fever group rickettsia from 514 samples from the 13 different tick species generated rickettsial-specific amplicons in 10% (54) of total ticks tested. Only three tick species Rhipicephalus microplus, Hyalomma anatolicum, and H. dromedarii had evidence of infection with “Candidatus Rickettsia amblyommii” a result further verified using a rompB gene-specific quantitative PCR (qPCR) assay. The Hyalomma ticks also tested positive for the piroplasm, Theileria annulata, using a qPCR assay. Conclusions/Significance This study provides information about tick diversity in Pakistan, and pathogenic bacteria in different tick species. Our results showed evidence for Candidatus R. amblyommii infection in Rhipicephalus microplus, H. anatolicum, and H. dromedarii ticks, which also carried T. annulata

Effect of angiotensin-converting enzyme inhibitor and angiotensin receptor blocker initiation on organ support-free days in patients hospitalized with COVID-19

IMPORTANCE Overactivation of the renin-angiotensin system (RAS) may contribute to poor clinical outcomes in patients with COVID-19. Objective To determine whether angiotensin-converting enzyme (ACE) inhibitor or angiotensin receptor blocker (ARB) initiation improves outcomes in patients hospitalized for COVID-19. DESIGN, SETTING, AND PARTICIPANTS In an ongoing, adaptive platform randomized clinical trial, 721 critically ill and 58 non–critically ill hospitalized adults were randomized to receive an RAS inhibitor or control between March 16, 2021, and February 25, 2022, at 69 sites in 7 countries (final follow-up on June 1, 2022). INTERVENTIONS Patients were randomized to receive open-label initiation of an ACE inhibitor (n = 257), ARB (n = 248), ARB in combination with DMX-200 (a chemokine receptor-2 inhibitor; n = 10), or no RAS inhibitor (control; n = 264) for up to 10 days. MAIN OUTCOMES AND MEASURES The primary outcome was organ support–free days, a composite of hospital survival and days alive without cardiovascular or respiratory organ support through 21 days. The primary analysis was a bayesian cumulative logistic model. Odds ratios (ORs) greater than 1 represent improved outcomes. RESULTS On February 25, 2022, enrollment was discontinued due to safety concerns. Among 679 critically ill patients with available primary outcome data, the median age was 56 years and 239 participants (35.2%) were women. Median (IQR) organ support–free days among critically ill patients was 10 (–1 to 16) in the ACE inhibitor group (n = 231), 8 (–1 to 17) in the ARB group (n = 217), and 12 (0 to 17) in the control group (n = 231) (median adjusted odds ratios of 0.77 [95% bayesian credible interval, 0.58-1.06] for improvement for ACE inhibitor and 0.76 [95% credible interval, 0.56-1.05] for ARB compared with control). The posterior probabilities that ACE inhibitors and ARBs worsened organ support–free days compared with control were 94.9% and 95.4%, respectively. Hospital survival occurred in 166 of 231 critically ill participants (71.9%) in the ACE inhibitor group, 152 of 217 (70.0%) in the ARB group, and 182 of 231 (78.8%) in the control group (posterior probabilities that ACE inhibitor and ARB worsened hospital survival compared with control were 95.3% and 98.1%, respectively). CONCLUSIONS AND RELEVANCE In this trial, among critically ill adults with COVID-19, initiation of an ACE inhibitor or ARB did not improve, and likely worsened, clinical outcomes. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT0273570

Bacterial diversity at the genus level in ticks from livestock in Pakistan.

<p>Group 1, <i>Rhipicephalus microplus</i> from cows; Group 2, <i>R</i>. <i>turanicus</i> from goats; Group 3, <i>Haemaphysalis cornupunctata</i> from sheep; Group 4, <i>Ha</i>. <i>cornupunctata</i> from goats; Group 5, <i>Ha</i>. <i>kashmerensis</i> from goats; Group 6, <i>Ha</i>. <i>montgomeryi</i> from goats; Group 7, <i>Ha</i>. <i>montgomeryi</i> from buffaloes; Group 8, <i>Ha</i>. <i>montgomeryi</i> from cows; Group 9, <i>Ha</i>. <i>bispinosa</i> from goats; Group 10, <i>Ha</i>. <i>bispinosa</i> from buffaloes; Group 11, <i>Hyalomma anatolicum</i> from cows; Group 12, <i>Hy</i>. <i>anatolicum</i> from buffaloes; Group 13, <i>Hy</i>. <i>scupense</i> from goats; Group 14, <i>Hy</i>. <i>isaaci</i> from cows; Group 15, <i>Ornithodoros tholozani</i> from buffaloes. Less than 2% of genera were removed during graph preparation. <i>Haemaphysalis</i> is abbreviated to <i>Ha</i>. <i>Hyalomma</i> is abbreviated to <i>Hy</i>.</p

Tick species diversity infesting livestock of Pakistan.

<p>(A) Tick species prevalent in different geographic regions of Pakistan (B) Tick species prevalence in Pakistan. Species with less than 1% abundance are grouped as “others”; these include <i>Haemaphysalis kashmirensis</i> (0.98), <i>Hyalomma turanicum</i> (0.67), <i>Rhipicephalus sanguineus</i> (0.39), <i>Hae</i>. <i>sulcata</i> (0.34), <i>Hy</i>. <i>kumari</i> (0.10), <i>Hy</i>. <i>hussaini</i> (0.08), and <i>R</i>. <i>annulatus</i> (0.08). <i>Haemaphysalis</i> is abbreviated to <i>Hae</i>. <i>Hyalomma</i> is abbreviated to <i>Hy</i>. Ticks (~ 4000) were collected from different livestock and domestic animal hosts (cattle, buffalo, sheep, goat, camel, dog, cat and poultry) across the country.</p

Piroplasm detection and quantification of <i>Theileria annulata</i> in <i>Hyalomma</i> ticks from livestock in Pakistan.

<p>Piroplasm detection and quantification of <i>Theileria annulata</i> in <i>Hyalomma</i> ticks from livestock in Pakistan.</p

Ticks tested for SFG Rickettsia.

<p>Ticks tested for SFG Rickettsia.</p

A study of ticks and tick-borne livestock pathogens in Pakistan

As obligate blood-feeding arthropods, ticks transmit pathogens to humans and domestic animals more often than other arthropod vectors. Livestock farming plays a vital role in the rural economy of Pakistan, and tick infestation causes serious problems with it. However, research on tick species diversity and tick-borne pathogens has rarely been conducted in Pakistan. In this study, a systematic investigation of the tick species infesting livestock in different ecological regions of Pakistan was conducted to determine the microbiome and pathobiome diversity in the indigenous ticks.A total of 3,866 tick specimens were morphologically identified as 19 different tick species representing three important hard ticks, Rhipicephalus, Haemaphysalis and Hyalomma, and two soft ticks, Ornithodorus and Argas. The bacterial diversity across these tick species was assessed by bacterial 16S rRNA gene sequencing using a 454-sequencing platform on 10 of the different tick species infesting livestock. The notable genera detected include Ralstonia, Clostridium, Staphylococcus, Rickettsia, Lactococcus, Lactobacillus, Corynebacterium, Enterobacter, and Enterococcus. A survey of Spotted fever group rickettsia from 514 samples from the 13 different tick species generated rickettsial-specific amplicons in 10% (54) of total ticks tested. Only three tick species Rhipicephalus microplus, Hyalomma anatolicum, and H. dromedarii had evidence of infection with "Candidatus Rickettsia amblyommii" a result further verified using a rompB gene-specific quantitative PCR (qPCR) assay. The Hyalomma ticks also tested positive for the piroplasm, Theileria annulata, using a qPCR assay.This study provides information about tick diversity in Pakistan, and pathogenic bacteria in different tick species. Our results showed evidence for Candidatus R. amblyommii infection in Rhipicephalus microplus, H. anatolicum, and H. dromedarii ticks, which also carried T. annulata