Inhibiting Glucose Metabolism Results in Herpes Simplex Encephalitis.

This report evaluates how HSV enters the brain to cause herpes simplex encephalitis following infection at a peripheral site. We demonstrate that encephalitis regularly occurred when BALB/c mice were infected with HSV and treated daily with 2-deoxy-d-glucose (2DG), which inhibits glucose use via the glycolysis pathway. The outcome of infection in the trigeminal ganglion (TG), the site to which the virus spreads, replicates, and establishes latency, showed marked differences in viral and cellular events between treated and untreated animals. In control-untreated mice, the replicating virus was present only during early time points, whereas in 2DG recipients, replicating virus remained for the 9-d observation period. This outcome correlated with significantly reduced numbers of innate inflammatory cells as well as T cells in 2DG-treated animals. Moreover, T cells in the TG of treated animals were less activated and contained a smaller fraction of expressed IFN-γ production compared with untreated controls. The breakdown of latency was accelerated when cultures of TG cells taken from mice with established HSV latency were cultured in the presence of 2DG. Taken together, the results of both in vivo and in vitro investigations demonstrate that the overall effects of 2DG therapy impaired the protective effects of one or more inflammatory cell types in the TG that normally function to control productive infection and prevent spread of virus to the brain.

Supplementing the Diet with Sodium Propionate Suppresses the Severity of Viral Immuno-inflammatory Lesions.

This report evaluates a dietary manipulation approach to suppress the severity of ocular infections caused by herpes simplex virus infection. The virus causes chronic damage to the cornea that results from a T-cell-orchestrated inflammatory reaction to the infection. Lesion severity can be limited if cells with regulatory activity predominate over proinflammatory T cells and nonlymphoid inflammatory cells. In this report, we show that this outcome can be achieved by including the short-chain fatty acid (SCFA) salt sodium propionate (SP) in the drinking water. Animals given the SP supplement developed significantly fewer ocular lesions than those receiving no supplement. Corneas and lymphoid organs contained fewer CD4 Th1 and Th17 T cells, neutrophils, and macrophages than those of controls, but a higher frequency of regulatory T cells (Treg) was present. The inclusion of SP in cultures to induce CD4 T cell subsets in vitro reduced the magnitude of Th1 and Th17 responses but expanded Treg induction. Dietary manipulation was an effective approach to limit the severity of viral immuno-inflammatory lesions and may be worth exploring as a means to reduce the impact of herpetic lesions in humans.IMPORTANCE Herpetic lesions are a significant problem, and they are difficult to control with therapeutics. Our studies show that the severity of herpetic lesions in a mouse model can be diminished by changing the diet to include increased levels of SCFA, which act to inhibit the involvement of inflammatory T cells. We suggest that changing the diet to include higher levels of SCFA might be a useful approach to reducing the impact of recurrent herpetic lesions in humans.

Modulating glutamine metabolism to control viral immuno-inflammatory lesions.

Infection of the cornea with HSV results in an immune-inflammatory reaction orchestrated by proinflammatory T cells that is a major cause of human vision impairment. The severity of lesions can be reduced if the representation of inflammatory T cells is changed to increase the presence of T cells with regulatory function. This report shows that inhibiting glutamine metabolism using 6-Diazo-5-oxo-l-norleucine (DON) administered via intraperitoneal (IP) starting 6 days after ocular infection and continued until day 15 significantly reduced the severity of herpetic stromal keratitis lesions. The therapy resulted in reduced neutrophils, macrophages as well proinflammatory CD4 Th1 and Th17 T cells in the cornea, but had no effect on levels of regulatory T cells. A similar change in the representation of inflammatory and regulatory T cells occurred in the trigeminal ganglion (TG) the site where HSV infection establishes latency. Glutamine metabolism was shown to be required for the in-vitro optimal induction of both Th1 and Th17 T cells but not for the induction of Treg that were increased when glutamine metabolism was inhibited. Inhibiting glutamine metabolism also changed the ability of latently infected TG cells from animals previously infected with HSV to reactivate and produce infectious virus.

Theileria equi and Babesia caballi infection of equids in Punjab, India: a serological and molecular survey.

A cross-sectional study was conducted in Submountain undulating, Undulating plain, Western and Western plain agro-climatic zones of Punjab province, India, to determine the prevalence, agreement between diagnostic tests and associated related risk factors of Theileria equi and Babesia caballi infection in equids (horses, donkey, mules). An overall prevalence of 14.14 and 0.0% of T. equi and B. caballi was recorded by multiplex polymerase chain reaction targeting 18S ribosomal RNA (rRNA) for both the parasites and 75 and 1.11% by competitive enzyme-linked immunosorbent assay in a representative sample of 180 animals. Only two animals with positive antibody titre from B. caballi and none with PCR indicated T. equi as the predominant haemoprotozoan responsible for equine piroplasmosis in the study area. Among the PCR-positive animals, presence of tick vectors in farm vicinity was the most influential associated with T. equi infection (P = 0.002; odds ratio (OR) 9.30; 95% confidence interval (CI) = 3.32-27.10). For animals with higher anti-T. equi antibody titres, strong association of sero-prevalence for T. equi was recorded with age, sex, usage, tick infestation and deworming/vaccination status of host animals and farm management strategies. The study has demonstrated the possible absence of B. caballi in both conducive and non-conducive areas of Punjab and demonstrated T. equi as the potential agent of equine piroplasmosis in Punjab.

Prevalence of haemoparasites infection in diseased bovine and canine of Bhatinda district of Punjab, India.

Infections caused by tick-borne haemoparasites pose a significant global threat to both human and animal health. Within this category, various haemoparasites species belonging to genera like Anaplasma sp., Babesia sp., Ehrlichia sp., Hepatozoon sp., and Theileria sp., are particularly concerning due to their ability to cause diseases in a wide range of hosts, including mammals, birds, and reptiles. The present cross-sectional study involving 580 animals provides annual insights into the prevalence of major haemoparasites infections in the Bathinda region of Punjab. The observed trends indicate that haemoparasites infections were most common in cattle, followed by buffalo and canines. Risk factor analysis revealed that crossbreed cattle were more susceptible to infection, with a prevalence of 35.73% (95% CI 4.28-45.17). Amongst the cattle, adults exhibited a higher vulnerability to haemoparasites infections, with a prevalence of 35.89% (95% CI 5.50-33.64). Conversely, companion animals showed the opposite pattern, with a prevalence of 18.18% (95% CI 9.11-169.27). Furthermore, female dogs had a higher risk of haemoparasites infection, with a prevalence of 16.28% (95% CI 8.36-218.7). In light of these findings, it is imperative to emphasize early diagnosis, prompt antiprotozoals drug treatment, and effective control of tick vectors for the successful recovery of animals afflicted by haemoparasites infections.

A metabolic blueprint of COVID-19 and long-term vaccine efficacy.

Viruses are obligatory protein-coated units and often utilize the metabolic functions of the cells they infect. Viruses hijack cellular metabolic functions and cause consequences that can range from minor to devastating, as we have all witnessed during the COVID-19 pandemic. For understanding the virus-driven pathogenesis and its implications on the host, the cellular metabolism needs to be elucidated. How SARS-CoV-2 triggers metabolic functions and rewires the metabolism remains unidentified but the implications of the metabolic patterns are under investigation by several researchers. In this review, we have described the SARS-CoV-2-mediated metabolic alterations from in vitro studies to metabolic changes reported in victims of COVID-19. We have also discussed potential therapeutic targets to diminish the viral infection and suppress the inflammatory response, with respect to evidenced studies based on COVID-19 research. Finally, we aimed to explain how we could extend vaccine-induced immunity in people by targeting the immunometabolism.

Controlling the Burden of COVID-19 by Manipulating Host Metabolism.

The coronavirus disease 2019 (COVID-19) pandemic caused by the coronavirus-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to cause global health problems, but its impact would be minimized if the many effective vaccines that have been developed were available and in widespread use by all societies. This ideal situation is not occurring so other means of controlling COVID-19 are needed. In this short review, we make the case that manipulating host metabolic pathways could be a therapeutic approach worth exploring. The rationale for such an approach comes from the fact that viruses cause metabolic changes in cells they infect, effective host defense mechanisms against viruses requires the activity of one or more metabolic pathways, and that hosts with metabolic defects such as diabetes are more susceptible to severe consequences after COVID-19. We describe the types of approaches that could be used to redirect various aspects of host metabolism and the success that some of these maneuvers have had at controlling other virus infections. Manipulating metabolic activities to control the outcome of COVID-19 has to date received minimal attention. Manipulating host metabolism will never replace vaccines to control COVID-19 but could be used as an adjunct therapy to the extent of ongoing infection.

Meta-analysis and comprehensive study of coronavirus outbreaks: SARS, MERS and COVID-19.

BACKGROUND: Zoonotic coronaviruses have caused several endemic and pandemic situations around the world. SARS caused the first epidemic alert at the beginning of this century, followed by MERS. COVID-19 appeared to be highly contagious, with human-to-human transmission by aerosol droplets, and reached nearly all countries around the world. A plethora of studies were performed, with reports being published within a short period of time by scientists and medical physicians. It has been difficult to find the relevant data to create an overview of the situation according to studies from accumulated findings and reports. In the present study we aimed to perform a comprehensive study in the context of the case fatality ratios (CFRs) of three major human Coronavirus outbreaks which occurred during the first twenty years of 21st century. METHODS: In this study, we performed meta-analyses on SARS, MERS and COVID-19 outbreak events from publicly available records. Study analyses were performed with the help of highly reputable scientific databases such as PubMed, WOS and Scopus to evaluate and present current knowledge on zoonotic coronavirus outbreaks, starting from 2000 to the end of 2020. RESULTS: A total of 250,194 research studies and records were identified with specific keywords and synonyms for the three viruses in order to cover all publications. In the end, 41 records were selected and included after applying several exclusion and inclusion criteria on identified datasets. SARS was found to have a nearly 11% case fatality ratio (CFR), which means the estimated number of deaths as a proportion of confirmed positive cases; Taiwan was the country most affected by the SARS outbreak based on the CFR analysis. MERS had CFRs of 35.8 and 26 in Saudi Arabia during the 2012 and 2015 outbreaks, respectively. COVID-19 resulted in a 2.2 CFR globally, and the USA reported the highest mortality ratio in the world in the end of first year of COVID-19 pandemic. CONCLUSION: Some members of the Coronaviridae family can cause highly contagious and devastating infections among humans. Within the last two decades, the whole world has witnessed several deadly emerging infectious diseases, which are most commonly zoonotic in nature. We conclude that pre-existing immunity during the early stages of a pandemic might be important, but case control and management strategies should be improved to decrease CFRs. Finally, we have addressed several concerns in relation to outbreak events in this study.

Could targeting immunometabolism be a way to control the burden of COVID-19 infection?

This review portrays the metabolic consequences of Covid-19 infection at different stages of the clinical syndrome. It also describes how events can change when patients with metabolic problems are infected and the effects that diet and nutrition might play to influence the outcome of infection. We also discuss the types of maneuvers that could be used to reshape metabolic events and question if this approach could be a practical therapy used alone or in combination with other approaches to reduce the burden of Covid-19 infection.

Virus Infections and Host Metabolism-Can We Manage the Interactions?

When viruses infect cells, they almost invariably cause metabolic changes in the infected cell as well as in several host cell types that react to the infection. Such metabolic changes provide potential targets for therapeutic approaches that could reduce the impact of infection. Several examples are discussed in this review, which include effects on energy metabolism, glutaminolysis and fatty acid metabolism. The response of the immune system also involves metabolic changes and manipulating these may change the outcome of infection. This could include changing the status of herpesviruses infections from productive to latency. The consequences of viral infections which include coronavirus disease 2019 (COVID-19), may also differ in patients with metabolic problems, such as diabetes mellitus (DM), obesity, and endocrine diseases. Nutrition status may also affect the pattern of events following viral infection and examples that impact on the pattern of human and experimental animal viral diseases and the mechanisms involved are discussed. Finally, we discuss the so far few published reports that have manipulated metabolic events in-vivo to change the outcome of virus infection. The topic is expected to expand in relevance as an approach used alone or in combination with other therapies to shape the nature of virus induced diseases.

Factors Affecting the Tissue Damaging Consequences of Viral Infections.

Humans and animals are infected by multiple endogenous and exogenous viruses but few agents cause overt tissue damage. We review the circumstances which favor overt disease expression. These can include intrinsic virulence of the agent, new agents acquired from heterologous species, the circumstances of infection such as dose and route, current infection with other agents which includes the composition of the microbiome at mucosal and other sites, past history of exposure to other infections as well as the immune status of the host. We also briefly discuss promising therapeutic strategies that can expand immune response patterns that minimize tissue damaging responses to viral infections.

Spatial seroepidemiology, risk assessment and haemato-biochemical implications of bovine trypanosomiasis in low lying areas of Punjab, India.

A cross-sectional study was carried out on 594 bovines (341 buffalo adults, 31 buffalo calves, 163 cattle adults, and 59 cattle calves) to assess the exposure of native bovine population to T. evansi elicited trypanosomiasis in the low-lying areas of Punjab (India). We ruled out the endemicity of the disease with 10.77% (95%CI = 8.53-13.52) sero-positive and 23.56% (95%CI = 20.33-27.15) suspected cases by card agglutination assay. We have presented the spatial distribution of these cases as a guideline to local veterinary practitioners and policy-makers. The categorical assessment of risk factors revealed buffalo adults are the most susceptible group in the state despite insignificant differences in farm management practices. A significant increase in the WBC, platelet, AST and serum iron, and decrease in haemoglobin, haematocrit volume, and serum glucose was recorded in both T. evansi positive and suspected animals.

Sero-prevalence and risk factor analysis of Theileria equi infection in equids from different agro-climatic zones of Punjab (India) by Indirect Immunofluorescence Antibody test.

In the Indian sub-continent, equine piroplasmosis (EP) is an imperative tick-borne disease of equids predominately caused by obligatory intra-erythrocytic protozoa Theileria equi. Present exploration is the first epidemiological survey report based on Indirect Immunofluorescence Antibody test (IFAT). A total of 120 equines (98 horses and 22 donkeys/mules) from five districts of two main agro-climatic zones of Punjab were screened. The overall prevalence was 58.33%, with western plane zone at higher infection risk for T. equi (74.50%, OR = 7.45, 95% CI = 62.24-85.76); followed by western zone (46.38%, OR = 4.63, 95% CI = 34-57.7). District wise highest and lowest prevalence was recorded in district Fazilka (80.76%) and Bathinda (35.29%) respectively. IFAT revealed elevated risk of infection of T. equi in the farms with inappropriate management structure. The risk factor connected with the type of host species had an odds ratio of 1.67 for donkeys/mules versus horses for T. equi infection. Followed by presence/absence of tick on animal. This study reveals the endemicity of EP in Punjab with prominence of various odds of infection, associated with latent infection of T. equi detectable by serological techniques as it was noticed that western plane zone and western zone are more prone to EP infection.

Molecular survey in relation to risk factors and haemato-biochemical alteration in Theileria equi infection of equines in Punjab Province, India.

Equine piroplasmosis caused by Theileria equi, an apicomplexan intracellular haemoprotozoan parasite effects equids throughout the world. Current investigation is the first detailed epidemiological survey report based on conventional (blood smear) and primary polymerase chain reaction (PCR) for the detection of T. equi on 464 equids (426 horses and 38 donkeys/mules) in Punjab province of India. PCR targeting 18S rRNA gene of T. equi produced high fidelity 709bp amplification products with 100% concordance with blood films. The prevalence of infection was proportional (P≤0.01) to temperature and aridness of the regions, which turned out to be the most important physical factor pertaining to T. equi infection. Spatial distribution analysis revealed an increasing trend of disease prevalence from north-eastern to south-western region of Punjab by both the techniques. Haemato-biochemical analysis revealed significant decrease in total erythrocyte count and haemoglobin; and increase in mean corpuscular haemoglobin, serum globulin, glucose, creatinine, aspartate aminotransferase and bilirubin levels (P≤0.05). This study divulges the endemicity of equine theileriosis in Punjab with the prominence of various odds of infection, emphasising the pathophysiological manifestation associated with latent infection of T. equi detectable by PCR.

Critical analysis of vector-borne infections in dogs: Babesia vogeli, Babesia gibsoni, Ehrlichia canis and Hepatozoon canis in Punjab, India.

There are few published studies on various vector borne diseases of dogs in India and most depict clinical infection in dogs, diagnosed by observation of the haemopathogens in stained blood smears. This study provides the first report regarding molecular confirmation and ancestral relationship analysis of blood smears positive cases of assorted haemopathogens in Punjab province of India. On blood smear examination, haemopathogens were observed in 124 out of 778 (15.95%, 95% CI: 13.53- 18.68) blood smears. Further polymerase chain reactions (PCR) was used on bloods smear positive cases to validate the results. Out of 778 blood samples, Babesia gibsoni was most common parasite infecting dogs (15.04%, 95% CI: 12.7-17.72), followed by Ehrlichia canis (0.39%, 95% CI: 0.0-1.13), infection of Babesia vogeli and Hepatozoon canis was same (0.26%, 95% CI: 0.0-0.9). Among various risk factors studied (age, sex, season), prevalence of infection was non-significantly higher in 1-2 year of age group (19.88%, 95% CI: 14.45-26.71), regarding sex same prevalence was recorded (15.94%), and chances of infection was highest in pre-monsoon i.e. summer (18.26%, 95% CI: 14.49-22.76). Phylogenetic analysis revealed ancestral background of Ludhiana isolates of B. vogeli, B. gibsoni, H. canis, and E. canis with the isolates of Philippines, Mongolia and Tunisia.

Spatial distribution, risk factors and haemato-biochemical alterations associated with Theileria equi infected equids of Punjab (India) diagnosed by indirect ELISA and nested PCR.

Equine piroplasmosis is a febrile, tick-borne disease of equids predominately caused by obligatory intra-erythrocytic protozoa Theileria equi in the Indian sub-continent. A cross-sectional study was carried out on 464 equids (426 horses and 38 donkeys/mules) in Punjab, India to assess the level of exposure to equine piroplasmosis by 18S rRNA gene nested polymerase chain reaction (nPCR) and equine merozoite antigen-2 (EMA2) indirect-ELISA (enzyme linked immunosorbent assay), to investigate risk factors and haemato-biochemical alterations associated with the infection. The endemicity of the disease was confirmed by positive PCR amplification in 21.77% and positive antibody titers in 49.78% equid samples. There was a fair agreement between these two diagnostic techniques (Kappa coefficient=0.326). The spatial distribution analysis revealed an increasing trend of T. equi prevalence from north-eastern to south-western region of Punjab by both the techniques correspondingly, which proffered a direct relation with temperature and inverse with humidity variables. The relatively prominent risk factor associated with sero-positivity was the presence of other domestic animals in the herd, while the propensity of finding a positive PCR amplification was higher in donkeys/mules, animal kept at unorganised farm or those used for commercial purposes as compared to their counterparts. There was a significant increase in globulins, gamma glutamyl-transferase, total bilirubin, direct bilirubin, indirect bilirubin, glucose levels and decrease in total erythrocyte count, haemoglobin, packed cell volume by animals, which were revealed positive by nPCR (may or may not positive by indirect-ELISA) and increase in creatinine, total bilirubin, direct bilirubin, glucose and decrease in total erythrocytes count by animals, which were revealed positive by indirect-ELISA (alone). To our knowledge, this study, for the first time, brings out a comprehensive report on the status on spatial distribution of T. equi in Punjab (India) state, thoroughly investigated by molecular and serological techniques, evaluating various environmental and demographic risk factors along with the haemato-biochemical alterations in the exposed animals.

Multiplex PCR for detection of Trypanosoma evansi and Theileria equi in equids of Punjab, India.

Multiplex PCR for simultaneous detection of Trypanosoma evansi and Theileria equi in single-step reaction was optimized and employed on 108 equids (99 horses and 9 donkeys/mules) blood samples collected from two agro-climatic zones (Sub-mountain undulating zone and Undulating plain zone) of Punjab to evaluate the status of concurrent infection and associated risk factors. The amplification products of 257 and 709 bp targeting repetitive nucleotide sequence of variable surface glycoproteins of T. evansi and 18S rRNA gene of T. equi, respectively expressed high fidelity of the primer pairs with sequence homology to neighboring geographic isolates. The overall prevalence of T. evansi and T. equi was 3.7 and 1.85%, with Undulating plain zone at higher infection risk for T. equi (OR=3.24, 95% CI=0.28-83.65); and Sub-mountain undulating zone (OR=∞, 95% CI=0.25-∞) for T. evansi. Multiplex PCR revealed higher risk of infection of both T. equi (OR=6.75, 95% CI=0.58-175.38) and T. evansi (OR=2.11, 95% CI=0.05-80.36) in the farms with inappropriate management system. The risk factor associated with the type of host species had an odds ratio of 12.35 (95% CI=0.29-508.37) for donkeys/mules versus horses for T. evansi infection. This group was also at higher risk of infection with Odds ratio (OR) of 4 (95% CI=0.14-53.99) for T. equi. The current investigation brings out various commodities at risk of infection pertaining to equid trypanosomosis and theileriosis evaluated by a rapid and sensitive multiplex PCR assay.

Equine trypanosomosis in central and western Punjab: prevalence, haemato-biochemical response and associated risk factors.

The detection of Trypanosoma evansi in blood is intricate, primarily in chronic stage of infection, as the parasitaemia is often low and fluctuating. The climatic conditions of the target area of Punjab (a province of India with a total of 34,000 horses and ponies used for sports and transport) are conducive for the parasite propagation. The objective of present investigation was to assess the prevalence of T. evansi in central and western Punjab by PCR and card agglutination test (CATT/T. evansi) in relation to clinico-haematobiochemical alterations and risk factors associated with latent trypanosomosis. A total of 169 equine blood and serum samples tested by CATT/T. evansi revealed 16 cases positive, with 6.8% from central plain and 13.63% from western zone. To assess the specificity of serological test, PCR1 was performed using established primer pair TR3 5'-GCG CGG ATT CTT TGC AGA CGA-3' and TR4 5'-TGC AGA CAC TGG AAT GTT ACT-3' for T. evansi. PCR2 applied with primer pair RoTat1.2F: 5'-ATG TCA ACG ATG CCT GTT ACA TTA CGC AC-3' and RoTat1.2R: 5'-TAA ATA TCA CTG TCA AGA CCT GCT GCG G-3' to rule out the consensus between the finding of the two PCR assays and agglutination test for T. evansi, which displayed results in concordance with PCR1. PCR assays showed 1.92 and 1.51% positive samples from central plain and western zone, respectively. With respect to PCR assay, CATT/T. evansi showed 100% sensitivity and 92.1% specificity. Microscopy showed a very low prevalence rate of 0.59% with only one sample positive with teaming parasitaemia. Comparison between sexes revealed higher positivity in mares by the three tests (BSE: 0.95%, PCR: 2.88%, CATT/T. evansi: 14.42%). The haemato-biochemical factors were found to be altered in PCR positive cases, while the mean value of vital parameters lied in normal range in seropositive cases. The female horse (RR=0.0937, 95% CI=1.388-190.223%) population was found to be at the highest risk of seropositivity for T. evansi, particularly in the unorganized farms (RR=19.726, 95% CI=2.918-400.221%).