Evaluation of trace elements in forages and their effect on gastrointestinal parasite burden in grazing sheep.

This study was designed to evaluate the trace elements (minerals) in forages fed to sheep and their effect on gastrointestinal parasite burdens. The ultimate objective was to determine the correlation between the burden of gastrointestinal (GI) parasites and the level of trace minerals in sheep serum as a result of the forages they grazed on. A total of 384 faecal samples were collected from sheep in each of the districts (Sialkot and Multan) and examined quantitatively using the McMaster technique. Serum collected from them and plants were pre-treated, and spectrophotometry was used to determine the concentration of trace minerals (Mn, Co, Cu, and Zn). The level of these trace elements differed significantly (P < 0.05) in forages from both districts. In the district of Sialkot, the highest concentrations (mg/Kg) of Zn (38.53 ± 0.16) were found in Cichorium intybus, Cu (41.57 ± 0.07) in Cynodon dactylon, Mn (39.61 ± 0.05) in Parthenium hysterophorus, and Co (1.42 ± 0.03) in Coronopus didymus. In the district of Multan, the highest concentrations (mg/Kg) of Zn (39.43 ± 0.46) were found in Cichorium intybus, Cu (25.76 ± 0.36) in Cynodon dactylon, Mn (34.29 ± 0.53) in Launaea nudicaulis, and Co (1.74 ± 0.08) in Brachiaria raptens. The prevalence of GI parasites in sheep populations in district Sialkot was 34%, while in district Multan, it was 32%. In tehsil Sialkot of district Sialkot, Zn and Cu were significantly (P < 0.05) correlated with eggs per gram (EPG) of faeces, while in tehsil Multan City of district Multan, only Cu was significantly (P < 0.05) correlated with EPG. The potential mechanism behind the role of trace minerals in lowering the burdens of GI parasites requires more investigation. It is recommended that plants with high content of trace minerals should be utilized as part of comprehensive preventive and control strategies against GI parasitism in ruminant animals like sheep.

Seasonal changes in the scrotal skin histology in relation to thermoregulation and testosterone level in camel and buffalo bull.

This study was conducted to compare the scrotal skin dynamics of camel and buffalo bulls regarding thermoregulation, season and testosterone. Scrotal skin and blood samples (n = 8 each) were collected during breeding (October to March) and non-breeding seasons (April to August). Tissue slides were prepared by the paraffin embedding technique followed by hematoxylin and eosin (H&E) staining and were analysed by ImageJ®. Relative humidity and temperature were recorded in different seasons to calculate the thermal humidity index (THI). Serum testosterone level was estimated through radioimmunoassay (RIA). The data were analysed by applying ANOVA, the THS test was used as a post-ANOVA interface and the correlation coefficient was also calculated among season, testosterone and skin parameters. The results showed that skin thickness, season and THI were negatively correlated with testosterone levels. In the neck, body and apex, the papillary layer was thicker in buffalo bull compared to camel bull during the breeding season; however, the reticular layer followed a reverse trend in the apex when THI was low. In the scrotal neck during the non-breeding season, hair follicle (HF) length and density were recorded high in camel bull compared to buffalo bull when THI was elevated. The density of HF followed the otherwise trend in the apex during the breeding season. The density of SG was found highest in all regions of camel bull as compared to buffalo bull and showed direct relation with THI. To conclude, this study delineates how the histo-dynamics of scrotal skin varies and accommodate itself with testosterone and THI over the different season in camel and buffalo.

Rabies Virus Exploits Cytoskeleton Network to Cause Early Disease Progression and Cellular Dysfunction.

Rabies virus (RABV) is a cunning neurotropic pathogen and causes top priority neglected tropical diseases in the developing world. The genome of RABV consists of nucleoprotein (N), phosphoprotein (P), matrix protein (M), glycoprotein (G), and RNA polymerase L protein (L), respectively. The virus causes neuronal dysfunction instead of neuronal cell death by deregulating the polymerization of the actin and microtubule cytoskeleton and subverts the associated binding and motor proteins for efficient viral progression. These binding proteins mainly maintain neuronal structure, morphology, synaptic integrity, and complex neurophysiological pathways. However, much of the exact mechanism of the viral-cytoskeleton interaction is yet unclear because several binding proteins of the actin-microtubule cytoskeleton are involved in multifaceted pathways to influence the retrograde and anterograde axonal transport of RABV. In this review, all the available scientific results regarding cytoskeleton elements and their possible interactions with RABV have been collected through systematic methodology, and thereby interpreted to explain sneaky features of RABV. The aim is to envisage the pathogenesis of RABV to understand further steps of RABV progression inside the cells. RABV interacts in a number of ways with the cell cytoskeleton to produce degenerative changes in the biochemical and neuropathological trails of neurons and other cell types. Briefly, RABV changes the gene expression of essential cytoskeleton related proteins, depolymerizes actin and microtubules, coordinates the synthesis of inclusion bodies, manipulates microtubules and associated motors proteins, and uses actin for clathrin-mediated entry in different cells. Most importantly, the P is the most intricate protein of RABV that performs complex functions. It artfully operates the dynein motor protein along the tracks of microtubules to assist the replication, transcription, and transport of RABV until its egress from the cell. New remedial insights at subcellular levels are needed to counteract the destabilization of the cytoskeleton under RABV infection to stop its life cycle.

Potential Therapeutic Benefits of Honey in Neurological Disorders: The Role of Polyphenols.

Honey is the principal premier product of beekeeping familiar to Homo for centuries. In every geological era and culture, evidence can be traced to the potential usefulness of honey in several ailments. With the advent of recent scientific approaches, honey has been proclaimed as a potent complementary and alternative medicine for the management and treatment of several maladies including various neurological disorders such as Alzheimer's disease, Parkinson's disease, Huntington's disease, and multiple sclerosis, etc. In the literature archive, oxidative stress and the deprivation of antioxidants are believed to be the paramount cause of many of these neuropathies. Since different types of honey are abundant with certain antioxidants, primarily in the form of diverse polyphenols, honey is undoubtedly a strong pharmaceutic candidate against multiple neurological diseases. In this review, we have indexed and comprehended the involved mechanisms of various constituent polyphenols including different phenolic acids, flavonoids, and other phytochemicals that manifest multiple antioxidant effects in various neurological disorders. All these mechanistic interpretations of the nutritious components of honey explain and justify the potential recommendation of sweet nectar in ameliorating the burden of neurological disorders that have significantly increased across the world in the last few decades.

Reprogramming Mycobacterium tuberculosis CRISPR System for Gene Editing and Genome-wide RNA Interference Screening.

Mycobacterium tuberculosis is the causative agent of tuberculosis (TB), which is still the leading cause of mortality from a single infectious disease worldwide. The development of novel anti-TB drugs and vaccines is severely hampered by the complicated and time-consuming genetic manipulation techniques for M. tuberculosis. Here, we harnessed an endogenous type III-A CRISPR/Cas10 system of M. tuberculosis for efficient gene editing and RNA interference (RNAi). This simple and easy method only needs to transform a single mini-CRISPR array plasmid, thus avoiding the introduction of exogenous protein and minimizing proteotoxicity. We demonstrated that M. tuberculosis genes can be efficiently and specifically knocked in/out by this system as confirmed by DNA high-throughput sequencing. This system was further applied to single- and multiple-gene RNAi. Moreover, we successfully performed genome-wide RNAi screening to identify M. tuberculosis genes regulating in vitro and intracellular growth. This system can be extensively used for exploring the functional genomics of M. tuberculosis and facilitate the development of novel anti-TB drugs and vaccines.

Concurrent Trypanosomosis, Babesiosis and Toxocarosis in a Dog: a Case Report.

Background: Among the blood-borne parasitic infections of dogs, trypanosomosis and babesiosis are highly prevalent in the Indian subcontinent and highly pathogenic. Methods: A six-month-old dog was presented with complaint of lethargy, anorexia and loss of body weight over the previous 15 days on October 11, 2021. A full clinical and targeted haematologic and parasitological examination was undertaken. Results: Wet smears were positive for the motile trypanosomes while stained thin blood smears showed large number of extracellular trypanosomes and intra-erythrocytic ring-like inclusion bodies of Babesia spp. A high burden of Toxocara canis was identified on coprological examination. Conclusion: This report describes the clinical manifestation of concurrent infection of haemoparasites and gastrointestinal nematodes in Pakistani dogs, and highlights the importance of prompt veterinary intervention.

Exploring rabies endemicity in Pakistan: Major constraints & possible solutions.

Rabies, the oldest recorded viral zoonosis in the Indo-Pakistan subcontinent, is a neglected and lingering endemic disease in Pakistan. The review of online available rabies-related journals, papers and reports through platforms and electronic databases have provided the basis for a detailed analysis of the rabies situation in Pakistan. Only published materials related to various research areas of rabies in Pakistan were included and meaningful conclusions were developed to identify major constraints and generate an intellectual discussion on possible solutions. Results revealed 52 studies representing major issues concerning rabies prevention related to topics including, but not limited to: epidemiological investigations (40.38%), prophylactic measures (21.15%), population of wandering dogs (13.46%), public awareness and government interventions (17.30%) and diagnostic surveillance (7.69%). In order to minimize these problems and reduce the prevalence of dog bites or rabies in significant manners, the country direly needs to apply the following actions: a maintained supply of rabies prophylactic measures in public hospitals at subsidized rates, mass dog vaccination at regional levels, enforced responsible animal ownerships, implementing a systematic One Health approach, and diagnostic labs equipped with surveillance mechanisms established in coordination with the livestock and medical departments. This review, which presents up-to-date information on the risk factors and epidemiological features of rabies in Pakistan, provides useful information for scientists, policy makers, and administrative health officials wishing to understand how this deadly disease persists in the absence of effective control measures.

Stealth Strategies of Mycobacterium tuberculosis for Immune Evasion.

Tuberculosis is a devastating disease causing high mortality all over the world, especially in the developing countries. Mycobacterium tuberculosis (M. tb) is the causative agent of tuberculosis which replicates in the intracellular environment of host macrophages. Although the host immune system is capable of completely eliminating the pathogen, co-evolution of M. tb with humans has resulted in its ability to hijack the host innate and adaptive immune systems in numerous ways. Limited recent progress has been made in the understanding of M. tb immune escape mechanisms, hence exploration of survival strategies of M. tb has been critically reviewed with an insight into understanding its pathogenesis. We summarized the recent studies regarding the modulation of innate immune response, adaptive immune response, epigenetics and the role of miRNA. All of these advancements suggest that M. tb is well-familiarize with the host immune system and possess the ability to hijack it for intracellular survival.

Detection of West Nile virus lineage 1 sequences in blood donors, Punjab Province, Pakistan.

OBJECTIVES: This study was performed to determine the presence of West Nile virus (WNV) in mosquito specimens and human blood donors in Pakistan. METHODS: A total of 4150 mosquito specimens were collected using CO2-baited traps from five selected districts of Punjab Province, Pakistan. The mosquitoes were taxonomically identified using standard morphological keys, resulting in 166 pools. In addition, 1070 serum samples were collected from human blood donors. RNA was extracted from mosquito and human samples and screened for WNV using a reverse transcriptase PCR (RT-PCR). RESULTS: None of the mosquito pools tested positive for WNV, whereas three samples from asymptomatic humans tested positive. To determine the WNV strains, partial sequences were compared against a global representation of 23 WNV sequences. The study strains were determined to come from WNV lineage 1. CONCLUSIONS: This study is novel in reporting the circulation of lineage 1 WNV in Pakistan. Given its ability to transmit from human to human via blood transfusion, this highlights the urgent need for nationwide surveillance to assess the distribution and impact of WNV in Pakistan. Determining the source of human infection will require more extensive mosquito sampling.

Transcriptional Study Revealed That Boron Supplementation May Alter the Immune-Related Genes Through MAPK Signaling in Ostrich Chick Thymus.

The objective of this study is to construct a digital gene expression tag profile to identify genes potentially related to immune response in the ostrich. Exposure to boron leads to an immune response in the ostrich, although the underlying mechanism remains obscure. Thus, a dire need of biological resource in the form of transcriptomic data for ostriches arises to key out genes and to gain insights into the function of boron on the immune response of thymus. For this purpose, RNA-Seq analysis was performed using the Illumina technique to investigate differentially expressed genes in ostrich thymuses treated with different boric acid concentrations (0, 80, and 640 mg/L). Compared with the control group, we identified 309 upregulated and 593 downregulated genes in the 80 mg/L treated sample and 228 upregulated and 1816 downregulated genes in 640 mg/L treated sample, respectively. Trend analysis of these differentially expressed genes uncovers three statistically significant trends. Functional annotation analysis of the differentially expressed genes verifies multiple functions associated with immune response. When ostrich thymuses were treated with boron, expression changes were observed in genes predominantly associated with MAPK and calcium signaling pathways. The results of this study provide all-inclusive information on gene expression at the transcriptional level that further enhances our apprehension for the molecular mechanisms of boron on the ostrich immune system. The calcium and MAPK signaling pathways might play a pivotal role in regulating the immune response of boron-treated ostriches.

Countrywide Survey for MERS-Coronavirus Antibodies in Dromedaries and Humans in Pakistan.

Middle East Respiratory Syndrome Coronavirus (MERS-CoV) is a zoonotic pathogen capable of causing severe respiratory disease in humans. Although dromedary camels are considered as a major reservoir host, the MERS-CoV infection dynamics in camels are not fully understood. Through surveillance in Pakistan, nasal (n = 776) and serum (n = 1050) samples were collected from camels between November 2015 and February 2018. Samples were collected from animal markets, free-roaming herds and abattoirs. An in-house ELISA was developed to detect IgG against MERS-CoV. A total of 794 camels were found seropositive for MERS-CoV. Prevalence increased with the age and the highest seroprevalence was recorded in camels aged > 10 years (81.37%) followed by those aged 3.1-10 years (78.65%) and ≤ 3 years (58.19%). Higher prevalence was observed in female (78.13%) as compared to male (70.70%). Of the camel nasal swabs, 22 were found to be positive by RT-qPCR though with high Ct values. Moreover, 2,409 human serum samples were also collected from four provinces of Pakistan during 2016-2017. Among the sampled population, 840 humans were camel herders. Although we found a high rate of MERS-CoV antibody positive dromedaries (75.62%) in Pakistan, no neutralizing antibodies were detected in humans with and without contact to camels.

Pharmacological effects and potential therapeutic targets of DT-13.

DT-13 is an isolated compound from Dwarf lillytruf tuber and currently among active research drugs by National Natural Science foundation of China for its several potential effects. The drug has been reported for its multiple pharmacological actions however no thorough review studies are available on it. Our present study is highlighting the pros and cons of DT-13 focusing on its potential pharmacological actions, therapeutic utilization and further exploration for novel targets. The drug possesses very low toxicity profile, quick onset and long duration of action with slow elimination that combinely makes it favorable for the clinical studies. In vivo and in vitro studies show that the drug regulates multiple cellular functions for its several pharmacological effects including, anti-adhesive effects via regulation of tissue factor and transforming growth factor; anti-migratory effects through indirect regulation of NM-IIA in the tumor microenvironment, Tissue factor, down-regulation of CCR5-CCL5 axis and MMP-2/9 inhibition; anti-metastatic effects via regulation of MMPs and tissue factor; pro-apoptotic effects by modulation of endocytosis of EGF receptor; anti-angiogenic effects via regulation of HIF-1α,ERK, Akt signalling and autophagy inducing characteristics by regulating PI3K/Akt/mTOR signalling pathway. In addition to anti-tumor activities, DT-13 has significant anti-inflammatory, cardioprotective, hepatoprotective and immunomodulating effects. Pharmaceutical dosage form and targeted drug delivery system for DT-13 has not been established yet. Moreover, DT-13, has not been studied for its action on brain, colorectal, hepatic, pancreatic, prostate and blood cancers. Similarly the effects of drug on carbohydrate and glucose metabolism is another niche yet to be explored. In some traditional therapies, crude drug from the plant is used against diabetic and neurological disorders that are not reported in scientific literature, however due to profound effects of DT-13 on blood and cerebral ischemic disorders, it is reasonable to hypothesize that there could be an association of DT-13 that require further exploration.

Vildagliptin loaded triangular DNA nanospheres coated with eudragit for oral delivery and better glycemic control in type 2 diabetes mellitus.

Diabetes mellitus type 2 is a multidimensional disease associated with poor glycemic control through compromised sensitivity of pancreatic islet α and ß cells against glucose and dwindled secretion of insulin which is linked with the quantity of incretin hormones that are abridged by dipeptidyl peptidase-4 (DPP-4) in diseased condition. Vildagliptin (VG) inhibits DPP-4 therefore regulates the incretins that conversely maintains glycemic control. The safe reach and absorption of VG from intestine was dubious. Therefore we used Electrostatic Attraction Method to develop drug loaded DNA nanotechnology triangles coated by Eudragit (Eud) to make stable nanospheres of Vildagliptin (VG). We further analyzed the formulated nanospheres by AFM, XRD, DSC, SEM, TGA, ATR-FTIR and native PAGE. Additionally the efficacy of formulated nanospheres for drug release and glycemic control was assessed in Db/Db mouse. Our results showed that formulated nanospheres are smooth, spherical, stable and uniform in size ranging from 500 to 2000 nm with drug entrapment efficiency up to 95 ± 2% and extended drug release up to 15 ± 2 h. FTIR and DSC results confirmed the absence of VG-DNA-Eud interaction and XRD studies revealed a change in the crystalline status of the VG in nanospheres. Ex-vivo studies indicate that Eud-DNA-VG nanospheres effectively bypasses the acidic pH of the stomach and enhances glycemic control in Db/Db mouse without any risk of pancreatitis or pancreatic cancer. To the best of our knowledge, this is the first study conclusively reporting that VG loaded DNA Nano-architects coated with Eudragit are stable, safe and may improve therapeutic outcomes after oral delivery.