Infection In Experimental Animals Research Articles

Influenza A virus within-host evolution and positive selection in a densely sampled household cohort over three seasons.

While influenza A virus (IAV) antigenic drift has been documented globally, in experimental animal infections, and in immunocompromised hosts, positive selection has generally not been detected in acute infections. This is likely due to challenges in distinguishing selected rare mutations from sequencing error, a reliance on cross-sectional sampling, and/or the lack of formal tests of selection for individual sites. Here, we sequenced IAV populations from 346 serial, daily nasal swabs from 143 individuals collected over three influenza seasons in a household cohort. Viruses were sequenced in duplicate, and intrahost single nucleotide variants (iSNVs) were identified at a 0.5% frequency threshold. Within-host populations exhibited low diversity, with >75% mutations present at <2% frequency. Children (0-5 years) had marginally higher within-host evolutionary rates than adolescents (6-18 years) and adults (>18 years, 4.4 × 10-6 vs. 9.42 × 10-7 and 3.45 × 10-6, P < .001). Forty-five iSNVs had evidence of parallel evolution but were not over-represented in HA and NA. Several increased from minority to consensus level, with strong linkage among iSNVs across segments. A Wright-Fisher approximate Bayesian computational model identified positive selection at 23/256 loci (9%) in A(H3N2) specimens and 19/176 loci (11%) in A(H1N1)pdm09 specimens, and these were infrequently found in circulation. Overall, we found that within-host IAV populations were subject to genetic drift and purifying selection, with only subtle differences across seasons, subtypes, and age strata. Positive selection was rare and inconsistently detected.

Increased Virulence of Culicoides Midge Cell-Derived Bluetongue Virus in IFNAR Mice.

Bluetongue (BT) is a Culicoides midge-borne hemorrhagic disease affecting cervids and ruminant livestock species, resulting in significant economic losses from animal production and trade restrictions. Experimental animal infections using the α/β interferon receptor knockout IFNAR mouse model and susceptible target species are critical for understanding viral pathogenesis, virulence, and evaluating vaccines. However, conducting experimental vector-borne transmission studies with the vector itself are logistically difficult and experimentally problematic. Therefore, experimental infections are induced by hypodermic injection with virus typically derived from baby hamster kidney (BHK) cells. Unfortunately, for many U.S. BTV serotypes, it is difficult to replicate the severity of the disease seen in natural, midge-transmitted infections by injecting BHK-derived virus into target host animals. Using the IFNAR BTV murine model, we compared the virulence of traditional BHK cell-derived BTV-17 with C. sonorensis midge (W8) cell-derived BTV-17 to determine whether using cells of the transmission vector would provide an in vitro virulence aspect of vector-transmitted virus. At both low and high doses, mice inoculated with W8-BTV-17 had an earlier onset of viremia, earlier onset and peak of clinical signs, and significantly higher mortality compared to mice inoculated with BHK-BTV-17. Our results suggest using a Culicoides W8 cell-derived inoculum may provide an in vitro vector-enhanced infection to more closely represent disease levels seen in natural midge-transmitted infections while avoiding the logistical and experimental complexity of working with live midges.

Oropouche Virus (OROV) in Pregnancy: An Emerging Cause of Placental and Fetal Infection Associated with Stillbirth and Microcephaly following Vertical Transmission.

Oropouche virus (OROV) is an emerging arbovirus endemic in Latin America and the Caribbean that causes Oropouche fever, a febrile illness that clinically resembles some other arboviral infections. It is currently spreading through Brazil and surrounding countries, where, from 1 January to 1 August 2024, more than 8000 cases have been identified in Bolivia, Brazil, Columbia, and Peru and for the first time in Cuba. Travelers with Oropouche fever have been identified in the United States and Europe. A significant occurrence during this epidemic has been the report of pregnant women infected with OROV who have had miscarriages and stillborn fetuses with placental, umbilical blood and fetal somatic organ samples that were RT-PCR positive for OROV and negative for other arboviruses. In addition, there have been four cases of newborn infants having microcephaly, in which the cerebrospinal fluid tested positive for IgM antibodies to OROV and negative for other arboviruses. This communication examines the biology, epidemiology, and clinical features of OROV, summarizes the 2023-2024 Oropouche virus epidemic, and describes the reported cases of vertical transmission and congenital infection, fetal death, and microcephaly in pregnant women with Oropouche fever, addresses experimental animal infections and potential placental pathology findings of OROV, and reviews other bunyavirus agents that can cause vertical transmission. Recommendations are made for pregnant women travelling to the regions affected by the epidemic.

Influenza A virus within-host evolution and positive selection in a densely sampled household cohort over three seasons.

While influenza A virus (IAV) antigenic drift has been documented globally, in experimental animal infections, and in immunocompromised hosts, positive selection has generally not been detected in acute infections. This is likely due to challenges in distinguishing selected rare mutations from sequencing error, a reliance on cross-sectional sampling, and/or the lack of formal tests of selection for individual sites. Here, we sequenced IAV populations from 346 serial, daily nasal swabs from 143 individuals collected over three influenza seasons in a household cohort. Viruses were sequenced in duplicate, and intrahost single nucleotide variants (iSNV) were identified at a 0.5% frequency threshold. Within-host populations were subject to purifying selection with >75% mutations present at <2% frequency. Children (0-5 years) had marginally higher within-host evolutionary rates than adolescents (6-18 years) and adults (>18 years, 4.4×10-6 vs. 9.42×10-7 and 3.45×10-6, p <0.001). Forty-five iSNV had evidence of parallel evolution, but were not overrepresented in HA and NA. Several increased from minority to consensus level, with strong linkage among iSNV across segments. A Wright Fisher Approximate Bayesian Computational model identified positive selection at 23/256 loci (9%) in A(H3N2) specimens and 19/176 loci (11%) in A(H1N1)pdm09 specimens, and these were infrequently found in circulation. Overall, we found that within-host IAV populations were subject to purifying selection and genetic drift, with only subtle differences across seasons, subtypes, and age strata. Positive selection was rare and inconsistently detected.

Preventive efficacy of preparations of vesicles of external membranes of atoxygenic strains of &lt;i&gt;Vibrio cholerae О1&lt;/i&gt; serogroup

Purpose: To evaluate the effectiveness and expediency of using vesicles of the outer membranes of atoxygenic strains of Vibrio cholerae O1 serogroup for the prevention of experimental cholera. Materials and methods: atoxigenic strains of V. cholerae O1 El Tor 18950 (ctxAB-tcpA-) and 18780 (ctxAB-tcpA+) isolated from the water of surface reservoirs were used to obtain vesicles. The protective properties of the obtained drugs were evaluated using a model of generalized cholera in white mice and a model of an isolated loop of the small intestine of an adult rabbit. Results: it has been shown that vesicles of the outer membranes isolated from atoxygenic strains of V. cholerae have a protective effect and prevent the development of infection in experimental animals when infected with a virulent strain of cholera. The obtained drugs are most effective when administered parenterally twice, with an interval of seven days, preventing the death of all white mice taken in the experiment and the development of pathogenetic processes in the small intestine of adult rabbits. Summary: the data obtained indicates the formation of pronounced immune protection against the disease in animals, as well as the possibility of using these structures to create preventive anti-cholera drugs.

SARS-CoV-2 Serological Investigation of White-Tailed Deer in Northeastern Ohio.

Coronaviruses are known to cross species barriers, and spill over among animals, from animals to humans, and vice versa. SARS-CoV-2 emerged in humans in late 2019. It is now known to infect numerous animal species, including companion animals and captive wildlife species. Experimental infections in other animals have established that many species are susceptible to infection, with new ones still being identified. We have developed an enzyme-linked immunosorbent assay (ELISA) for detecting antibodies to SARS-CoV-2 nucleocapsid (N) and spike (S) proteins, that is both sensitive and specific. It can detect S antibodies in sera at dilutions greater than 1:10,000, and does not cross-react with antibodies to the other coronaviruses tested. We used the S antibody ELISA to test serum samples collected from 472 deer from ten sites in northeastern Ohio between November 2020 and March 2021, when the SARS-CoV-2 pandemic was first peaking in humans in Ohio, USA. Antibodies to SARS-CoV-2 were found in serum samples from every site, with an overall positivity rate of 17.2%; we further compared the viral neutralizing antibody titers to our ELISA results. These findings demonstrate the need to establish surveillance programs to monitor deer and other susceptible wildlife species globally.

A simple method for enrichment of phase I Coxiella burnetii

Coxiella burnetii is the bacterial causative agent of the zoonosis Q fever. This bacterium undergoes lipopolysaccharide (LPS) phase transition similar to Enterobacteriaciae upon in vitro passage. Full-length, phase I C. burnetii LPS is a critical virulence factor and profoundly impacts vaccine-induced immunogenicity; thus, LPS phase is an important consideration in C. burnetii experimentation and Q fever vaccine design. Typically, phase I LPS-expressing organisms are obtained from the tissues of infected experimental animals. In this process, residual phase II LPS-expressing organisms are thought to be cleared by the host immune system. Here, we propose an efficient and non-animal-based method for the enrichment of C. burnetii phase I LPS-expressing bacteria in vitro. We utilize both Vero cell culture to selectively enrich solutions with phase I and intermediate phase LPS-expressing bacteria. This simple and quick method decreases reliance on experimental animals and is a sustainable solution for Q fever diagnostic and vaccine development hurdles.

FcRγ- NK Cell Induction by Specific Cytomegalovirus and Expansion by Subclinical Viral Infections in Rhesus Macaques.

"Adaptive" NK cells, characterized by FcRγ deficiency and enhanced responsiveness to Ab-bound, virus-infected cells, have been found in certain hCMV-seropositive individuals. Because humans are exposed to numerous microbes and environmental agents, specific relationships between hCMV and FcRγ-deficient NK cells (also known as g-NK cells) have been challenging to define. Here, we show that a subgroup of rhesus CMV (RhCMV)-seropositive macaques possesses FcRγ-deficient NK cells that stably persist and display a phenotype resembling human FcRγ-deficient NK cells. Moreover, these macaque NK cells resembled human FcRγ-deficient NK cells with respect to functional characteristics, including enhanced responsiveness to RhCMV-infected target in an Ab-dependent manner and hyporesponsiveness to tumor and cytokine stimulation. These cells were not detected in specific pathogen-free (SPF) macaques free of RhCMV and six other viruses; however, experimental infection of SPF animals with RhCMV strain UCD59, but not RhCMV strain 68-1 or SIV, led to induction of FcRγ-deficient NK cells. In non-SPF macaques, coinfection by RhCMV with other common viruses was associated with higher frequencies of FcRγ-deficient NK cells. These results support a causal role for specific CMV strain(s) in the induction of FcRγ-deficient NK cells and suggest that coinfection by other viruses further expands this memory-like NK cell pool.

Influence of siRNA complexes on the reproduction of influenza A virus (Orthomyxoviridae: Alphainfluenzavirus) in vivo

Influenza is one of the most pressing global health problems. Despite the wide range of available anti-influenza drugs, the viral drug resistance is an increasing concern and requires the search for new approaches to overcome it. A promising solution is the development of drugs with action that is based on the inhibition of the activity of cellular genes through RNA interference. Evaluation in vivo of the preventive potential of miRNAs directed to the cellular genes FLT4, Nup98 and Nup205 against influenza infection. The A/California/7/09 strain of influenza virus (H1N1) and BALB/c mice were used in the study. The administration of siRNA and experimental infection of animals were performed intranasally. The results of the experiment were analyzed using molecular genetic and virological methods. The use of siRNA complexes Nup98.1 and Nup205.1 led to a significant decrease in viral reproduction and concentration of viral RNA on the 3rd day after infection. When two siRNA complexes (Nup98.1 and Nup205.1) were administered simultaneously, a significant decrease in viral titer and concentration of viral RNA was also noted compared with the control groups. The use of siRNAs in vivo can lead to an antiviral effect when the activity of single or several cellular genes is suppressed. The results indicate that the use of siRNAs targeting the cellular genes whose expression products are involved in viral reproduction is one of the promising methods for the prevention and treatment of not only influenza, but also other respiratory infections.

Hepatitis E as a Zoonosis.

Hepatitis E viruses in the family of Hepeviridae have been classified into 2 genus, 5 species, and 13 genotypes, involving different animal hosts of different habitats. Among all these genotypes, four (genotypes 3, 4, 7, and C1) of them are confirmed zoonotic causing sporadic human diseases, two (genotypes 5 and 8) were likely zoonotic showing experimental animal infections, and the other seven were not zoonotic or unconfirmed. These zoonotic HEV carrying hosts include pig, boar, deer, rabbit, camel, and rat. Taxonomically, all the zoonotic HEVs belong to the genus Orthohepevirus, which include genotypes 3, 4, 5, 7, 8 HEV in the species A and genotype C1 HEV in the species C. In the chapter, information of zoonotic HEV such as swine HEV (genotype 3 and 4), wild boar HEV (genotypes 3-6), rabbit HEV (genotype 3), camel HEV (genotype 7 and 8), and rat HEV (HEV-C1) was provided in detail. At the same time, their prevalence characteristics, transmission route, phylogenetic relationship, and detection technology were discussed. Other animal hosts of HEVs were introduced briefly in the chapter. All these information help peer researchers have basic understanding of zoonotic HEV and adopt reasonable strategy of surveillance and prevention.

Influence of Sitophilus oryzae on biological properties of Mycobacterium bovis

The active spread of tuberculosis around the world has been an urgent issue for many decades, but even now, there is no data on all available methods of introducing the pathogen into a previously healthy territory and the mechanisms of mycobacteria activity, and this makes it impossible to eradicate the disease completely. The purpose of this study was to determine the effect of the rice weevil (lat. Sitophilus oryzae) on the dissociative forms of mycobacterium tuberculosis, their viability, biochemical properties and virulence through passage of the pathogen through the insect’s body. To fulfil this purpose, the following methods were used: biological (experimental infection of laboratory animals), anatomical pathology and determining the viability of mycobacteria by counting colony-forming units. The effect on pathogenic properties of dissociative forms of Mycobacterium bovis (passage 118) after passage through the body of the rice weevil in an experiment on guinea pigs was determined and summarized. The change in the viability of mycobacteria isolated in the dynamics of the experiment in comparison with the original culture was described. The plasticity of the biochemical activity of the pathogen after passing through the body of the beetle was proven, and the influence of the macroorganism on the enzymatic properties of mycobacterium tuberculosis was analysed. The phenomena of the activation of enzymatic systems, which are associated with the adaptation and survival of mycobacteria in the conditions of a changed habitat, were substantiated. The practical value of this study lies in the improved understanding of the processes of the spread of mycobacterium tuberculosis and the established role of insects in the transmission of infection, with the prospect of developing measures to prevent and combat the disease

Pathogenicity of a novel classical swine fever LOM vaccine-derived virus isolated on Jeju Island, South Korea.

Reemergent local outbreaks of classical swine fever (CSF) occurred simultaneously in multiple pig farms on CSF-free Jeju Island, South Korea, in 2014 because of inadvertent injection of a commercial CSF (LOM) vaccine into pregnant sows. The LOM virus has since spread across the island and has become endemic in Jeju herds, raising concern about possible reversion to the virulence of the LOM vaccine. We previously isolated LOM-derived field CSF virus (CSFV) strains with unique insertion-deletion (INDEL) mutations in the 3'-untranslated region (UTR), designated LOM-derived Jeju 3'-UTR INDEL variants, from CSF-recurrent swine farms on Jeju Island in 2019. The present study conducted animal experiments to investigate whether a 2019 emergent LOM 3'-UTR INDEL variant, KNU-1905, has reverted to a pathogenic form in conventional pigs (n = 10). Experimental animal infection showed that pigs inoculated with the commercial LOM vaccine strain developed no adverse effects compared to the sham-infected pigs. However, KNU-1905 displayed pathogenic characteristics in pigs, including clinical symptoms (e.g., lethargy, conjunctivitis, nasal discharge, and diarrhoea), weight loss, and gross lesions. Moreover, viremia, virus shedding in faeces and nasal fluids, and viral loads in various tissues of all the KNU-1905-infected pigs were highly significant, in contrast to those of the LOM-infected group in which CSFV RNA was detected only in the serum, nasal, and tonsil samples of one identical pig. Overall, the LOM-derived field isolate with molecular variations induced clinical adverse events in pigs, which commonly shed considerable amounts of CSFV. This study provides evidence that the genetic evolution of the LOM-derived CSFV circulating on Jeju Island might have allowed the LOM vaccine to recover its primary prototype and that these variants might have induced chronic or persistent infection in pigs that can shed CSFV in field farms leading to a risk of transmission among pigs or farms in this former CSF-free region.

Morphological and Molecular Characterization, and Demonstration of a Definitive Host, for Sarcocystis masoni from an Alpaca (Vicugna pacos) in China.

Simple SummarySarcocystis spp. are cyst-forming intracellular protozoan parasites characterized by a two-host prey–predator life cycle. The alpaca (Vicugna pacos) is one of the South American camelids (SACs), and in recent years, this animal was introduced to China to be raised for its meat, skin, and wool and to be kept as tourist attractions and as pets. There is considerable confusion regarding the classification and nomenclature of the species of Sarcocystis in SACs. Two Sarcocystis species, named S. auchenia and S. masoni, are currently regarded as valid in SACs based on sarcocyst morphology and 18S rDNA sequences. However, the definitive host of S. masoni remains unknown. Here, S. masoni sarcocysts in an alpaca were morphologically described and molecularly characterized. Furthermore, the life cycle of S. masoni was completed via experimental animal infection. The present analysis showed that S. masoni has a close relationship with S. cameli in the dromedary camel (Camelus dromedaries), and the relationship between the two parasites needs to be clarified in the future.Only 18S rDNA sequences of Sarcocystis spp. in South American camelids (SACs) are deposited in GenBank as references, and the definitive host of S. masoni in SACs is still unclear. Here, S. masoni sarcocysts detected in an alpaca (Vicugna pacos) in China were investigated with the aid of light (LM) and transmission electron (TEM) microscopy, and characterized using four genetic markers, i.e., 18S rDNA, 28S rDNA and ITS, and the mitochondrial cox1. Additionally, the life cycle of the parasite was completed via experimental animal infection. Under LM, S. masoni sarcocysts exhibited numerous 1.3–2.1 μm conical protrusions. Under TEM, the sarcocyst wall contained conical, cylindrical, or irregular-shaped villar protrusions, similar to type 9j. Two dogs (Canis familiaris) fed S. masoni sarcocysts shed sporocysts with a prepatent period of 8–9 days. The newly obtained 18S rDNA sequences showed 98.4–100% identity with those of S. masoni in SACs previously deposited in GenBank. Interestingly, the newly obtained sequences of 18S rDNA and mitochondrial cox1 shared 99.6–100% and 98.2–98.5% identity, respectively, with those of S. cameli in dromedary camels (Camelus dromedaries). Phylogenetic analysis based on sequences of 18S rDNA, 28S rDNA, or mitochondrial cox1 revealed that S. masoni has a close relationship with Sarcocystis spp. in ruminants. The relationship between S. masoni and S. cameli deserves to be further clarified in the future.

Different panel of toll-like receptors expression during chronic Schistosoma mansoni infection in experimental animals

BackgroundMany studies have reported the immunomodulatory effect of helminths to avoid the lethal immunopathology. During schistosomiasis, the immune response is orchestrated by toll-like receptors (TLRs). Modulating TLRs can alter the function of antigen presentation cells with the shift of the host's Th1 response to a dominant regulatory Th2 response. The objective of our study was to clarify which TLRs are related to the immune response of chronic Schistosoma infection. MethodsThe study animals were divided into two groups; group I: uninfected mice; control group and group II: Schistosoma mansoni infected mice. mRNA expression of TLR2, 3, 4, 7, and 9 in different organs (liver, large intestine, and spleen) were assessed on day 90 post-infection. ResultsTLR gene expression has changed depending on the tissue studied as the mRNA level of TLR2, TLR7, and TLR9 were significantly upregulated in all examined organs while TLR3 expression showed only significant upregulation in the liver of infected mice. On the other hand, TLR4 expression was significantly upregulated in the liver while significantly downregulated in the large intestine. ConclusionThis study provides a better understanding of TLRs profile in different organs against S. mansoni parasites during the chronic phase of infection.

Evaluation of the efficacy of gold nanoparticles on Giardia lamblia infection in experimental animals

Giardia lamblia (G. lamblia) is an important cause of severe malabsorption, weight loss, physical and mental retardation especially in infants and children throughout the world. Metronidazole (MTZ) is the standard drug used for their treatment which possesses several drawbacks with low efficacy. Gold nanoparticles possess a broad-spectrum antimicrobial activity and could be considered as a future alternative to many microbial agents. This study aimed to evaluate the anti-Giardia effect of gold nanoparticles as an alternative to MTZ. This study was done on 70 experimentally albino rats that were divided into three main groups with seven subgroups (each of 10 rats). The effect of MTZ and gold nanoparticles as single or combined therapy were evaluated. The effect was assessed by counting Giardia fecal cysts in the stool and trophozoites in the intestinal wash, histopathological, transmission and scanning electron microscopic examinations of the small intestinal tissues. Toxic tests of biochemical parameters of liver and kidney function were also performed. A significant reduction of the parasite number in the stool and small intestinal sections was apparent in treated infected rats compared with the infected non-treated ones. Gold nanoparticles showed the best result and the highest effect in the eradication of the parasite from the stool and the intestine with marked improvement in the intestinal mucosal injury caused by G. lamblia trophozoites. Gold nanoparticles had a toxic effect on the liver, with no kidney toxicity. Nanogold can be considered as a potential therapeutic agent and as a promising alternative therapy for G. lamblia infection. Further studies using various dosages with different durations of treatment with gold nanoparticles can be tested on Giardia lamblia infection.

Anticoccidial effects of the Houttuynia cordata extract on experimental animal infection model

Coccidiosis is caused by infection of Eimeria species and an significant parasitic disease in poultry. Various kinds of natural products have been studied to find alternative treatments for coccidiosis in chickens, but the effect of Houttuynia cordata on Eimeria infection has not been investigated. The aim of this study is to study the anticoccidial effect of H. cordata extract (HCE) in chickens after oral infection by Eimeria tenella. Anticoccidial effects of the HCE was evaluated in chickens after oral infection with E. tenella. This study was performed on three-day-old chicks (n = 30). These animals were divided into 3 groups; HCE 0.2% treated/infected (n = 10), HCE untreated/infected (n = 10) and non-infected control (n = 10). The effect of HCE on E. tenella infection was assessed by two parameters; fecal oocysts shedding and body weights gain. the chicks fed HCE significantly reduced fecal oocysts when compared to the E. tenella-infected group fed standard diets (p&lt;0.05). Furthermore, the HCE-based diet improved weight loss due to E. tenella infection. Our data shows that HCE had significant antiprotozoal activity against E. tenella. These findings may have implications for the development of anticoccidial drugs.

Experimental investigations on effects of ZnO NPS and annona muricata extract for in vitro and in vivo antibacterial activity

ZnO NPs and annona muricata extract was used as antibacterial against some pathogenic bacteria isolated from different sources of infections. Furthermore, this work demonstrated the effect of oral dosage of ZnO NPs and annona muricata extract on the infected experimental animals. Moreover, the results showed the sensitivity of studied bacteria is variance against ZnO NPs and annona muricata extract and this sensitivity increase with increasing the concentration of both ZnO NPs and annona muricata extract. The in vitro study included using five bacterial species (S.aureus, S.epidermidis, P.aeruginosa, Proteus sp. and E.coli) and two in vivo studies (S.aureus and E.coli) which were given orally to experimental animals to verify the antibacterial activity of ZnO NPs and annona muricata extract against infections resulting from these pathogenic bacteria. The results were showed the effectiveness of ZnO NPs and annona muricata extract and no negative effect to be used as antibacterial agents. Therefore, this study can be considered important in the field of medicine, because the results of healing are very effective for these two substances.

Flow cytometry evaluation of the rhesus monkey cellular immunity following the Zaire ebolavirus (Filoviridae; Ebolavirus: Zaire ebolavirus) experimental infection

The outbreaks of the Zaire ebolavirus (ZE) disease (ZED) that have arisen in the last decade determine the need to study the infection pathogenesis, the formation of specific immunity forming as well as the development of effective preventive and therapeutic means. All stages of fight against the ZED spread require the experimental infection in sensitive laboratory animals, which are rhesus monkeys in case of this disease .The aim of the study is to evaluate the rhesus monkey cellular immunity following the ZE experimental infection by the means of flow cytometry (cytofluorimetry). Male rhesus monkeys were intramuscularly infected by the dose of 15 LD50 (dose of the pathogen that causes 50% mortality of infected animals) of the ZE, the Zaire strain (ZEBOV). Levels of 18 peripheral blood lymphocyte populations of the animals before the ZE experimental infection and at the terminal stage of the disease were assessed using flow cytometry. The certain changes in the levels of the lymphocyte populations were observed following infection, indicating simultaneous activation and suppression of the immune system during ZED. The increase in content was observed for T-lymphocytes, T-helper and cytotoxic T-lymphocytes expressing the corresponding markers of early activation. The decrease was recorded for T-lymphocytes and double-positive T-lymphocytes expressing corresponding markers of late activation, as well as natural killer cells expressing CD8 (p &lt; 0.05). For the first time in the Russian Federation, the rhesus monkey cellular immunity before and after the ZE experimental infection was assessed using flow cytometry.

Genomic characterization of 99 viruses from the bunyavirus families Nairoviridae, Peribunyaviridae, and Phenuiviridae, including 35 previously unsequenced viruses.

Bunyaviruses (Negarnaviricota: Bunyavirales) are a large and diverse group of viruses that include important human, veterinary, and plant pathogens. The rapid characterization of known and new emerging pathogens depends on the availability of comprehensive reference sequence databases that can be used to match unknowns, infer evolutionary relationships and pathogenic potential, and make response decisions in an evidence-based manner. In this study, we determined the coding-complete genome sequences of 99 bunyaviruses in the Centers for Disease Control and Prevention's Arbovirus Reference Collection, focusing on orthonairoviruses (family Nairoviridae), orthobunyaviruses (Peribunyaviridae), and phleboviruses (Phenuiviridae) that either completely or partially lacked genome sequences. These viruses had been collected over 66 years from 27 countries from vertebrates and arthropods representing 37 genera. Many of the viruses had been characterized serologically and through experimental infection of animals but were isolated in the pre-sequencing era. We took advantage of our unusually large sample size to systematically evaluate genomic characteristics of these viruses, including reassortment, and co-infection. We corroborated our findings using several independent molecular and virologic approaches, including Sanger sequencing of 197 genome segments, and plaque isolation of viruses from putative co-infected virus stocks. This study contributes to the described genetic diversity of bunyaviruses and will enhance the capacity to characterize emerging human pathogenic bunyaviruses.

Investigating chemical changes during Snake fruit and Black tea Kombucha fermentation and the associated immunomodulatory activity in Salmonella typhi-infected mice

This study uncovered the chemical changes during kombucha's fermentation process and revealed the associated immunomodulatory activity in Salmonella typhi-infected mice. The snake fruit juice and black tea extract were processed into kombucha (a beverage known for its health benefits) by fermentation with SCOBY culture at room temperature for 14 days. Snake fruit kombucha showed high changes in fermentation parameters (total acidity, pH, and total sugar), as well as bioactive compounds and antioxidant activity. Salmonella typhi demonstrated a reduction in the population of CD8+TNFα+ and CD4+IFNγ+ of infected experimental animals. Both snake fruit kombucha and black tea kombucha have the potential to be utilized as an immunomodulator to circumvent unstable conditions of the immune system caused by Salmonella typhi. Black tea kombucha and snake fruit kombucha cancan raise the production of CD8+TNFα+ and CD4+IFNγ+ in mice infected with Salmonella typhi. In the group of normal mice, black tea and snake fruit kombucha were able to lower down the production of CD8+TNFα+, which is a potent mechanism to modulate the immune system. Further research is required to highlight the mechanism and role of black tea kombucha and snake fruit kombucha in the immune response that modulates and treats infection by Salmonella typhi.