Contagious Pathogen Research Articles

Development of an accurate and rapid method for whole genome characterization of canine parvovirus

Canine parvovirus is a highly contagious pathogen affecting domestic dogs and other carnivores globally. Monitoring CPV through continuous genomic surveillance is crucial for mapping variability and developing effective control measures. Here, we developed a method using multiplex-PCR-next-generation sequencing to obtain full-length CPV genomes directly from clinical samples. This approach utilizes tiling and tailed amplicons to amplify overlapping fragments of roughly 250 base pairs. This enables the creation of Illumina libraries by conducting two PCR reaction runs. We tested the assay in 10 fecal samples from dogs diagnosed with CPV and one CPV-2 vaccine strain. Furthermore, we applied it to a feline sample previously diagnosed with the feline panleukopenia virus. The assay provided 100 % genome coverage and high sequencing depth across all 12 samples. It successfully provided the sequence of the coding regions and the left and right non-translated regions, including tandem and terminal repeats. The assay effectively amplified viral variants from divergent evolutionary groups, including the antigenic variants (2a, 2b, and 2c) and the ancestral CPV-2 strain included in vaccine formulations. Moreover, it successfully amplified the entire genome of the feline panleukopenia virus found in cat feces. This method is cost-effective, time-efficient, and does not require lab expertise in Illumina library preparation. The multiplex-PCR-next-generation methodology facilitates large-scale genomic sequencing, expanding the limited number of complete genomes currently available in databases and enabling real-time genomic surveillance. Furthermore, the method helps identify and track emerging CPV viral variants, facilitating molecular epidemiology and control. Adopting this approach can enhance our understanding of the evolution and genetic diversity of Protoparvovirus carnivoran1.

Innovative Elastomers with Antimicrobial Activity May Decrease Infection Risks during Milking.

Contagious pathogens are very costly to dairy herds, and they may have zoonotic and reverse-zoonotic potentials and may contribute to the spread of antimicrobial resistance. One of the most important risk factors for spreading these infections is milking, when liner contamination may transfer the pathogens from infected to healthy cows. There is no effective protocol to prevent the transmission of infection without the segregation of infected cows. Recently, the availability of elastomers with patented antimicrobial components in their formulations has allowed the exploration of alternative methods to reduce the risk of infection. Two different types of elastomers (rubber and silicone) and nine different formulations were challenged with three major mastitis pathogens (S. aureus, S. agalactiae, and E. coli). The results that were obtained in this study were interesting and unexpected. Indeed, to our knowledge, this is the first study to show that basic rubber materials have intrinsic antimicrobial activity. Silicone elastomers did not exhibit the same levels of bactericidal activity, although they did exhibit some antibacterial capacity. A significant decrease in bacterial survival curves was observed for all the formulations tested when antimicrobial components were added. The different results observed for the various products are likely due to the different formulations and diverse manufacturing processes. The availability of these new materials that significantly reduce the bacterial load on the liner surface may reduce the risk of spreading intramammary infections during milking. This would be an important step forward in achieving global sustainability of dairy herds, consistent with the objectives of One Health, by reducing the risks of zoonotic diseases and antimicrobial treatments.

Canine distemper virus (CDV)-neutralizing activities of an anti-CDV canine-derived single-chain variable antibody fragment 4-15 (scFv 4-15) screened by phage display technology

Canine distemper virus (CDV) is a highly contagious pathogen that causes severe diarrhea, fever and vomiting in domestic dogs, posing a serious threat to the dog breeding industry. Currently, there are no effective therapeutic agents for emergency treatment despite the availability of vaccines against CDV infection. Single-chain fragment variable (scFv) antibody has been demonstrated to effectively inhibit virus infections, suggesting a potential candidate as a therapeutic agent for canine distemper. In this study, a phage-displayed scFv library was constructed from the peripheral blood lymphocytes of dog immunized intramuscularly with live-attenuated CDV vaccine, and was subjected to four rounds of pannings against CDV. Subsequent indirect enzyme-linked immunosorbent assay screening revealed high-affinity scFv antibodies specific to CDV, and indirect immunofluorescence assay screening revealed CDV-neutralizing activity of scFv antibodies. Our results showed that a scFv antibody 4–15 (scFv 4–15) with high-affinity binding to CDV and neutralizing activity against CDV was obtained, which displayed effective therapeutic potential in vivo for dogs challenged with a lethal dose of CDV. Conclusively, the scFv 4–15 with high-affinity binding and neutralizing activity to CDV that was obtained by phage display technology provides a promising candidate for the therapeutic agents against CDV infection.

A multiplexed RT-PCR assay for nanopore whole genome sequencing of Tilapia lake virus (TiLV)

Tilapia lake virus (TiLV) is a highly contagious viral pathogen that affects tilapia, a globally significant and affordable source of fish protein. To prevent the introduction and spread of TiLV and its impact, there is an urgent need for increased surveillance, improved biosecurity measures, and continuous development of effective diagnostic and rapid sequencing methods. In this study, we have developed a multiplexed RT-PCR assay that can amplify all ten complete genomic segments of TiLV from various sources of isolation. The amplicons generated using this approach were immediately subjected to real-time sequencing on the Nanopore system. By using this approach, we have recovered and assembled 10 TiLV genomes from total RNA extracted from naturally TiLV-infected tilapia fish, concentrated tilapia rearing water, and cell culture. Our phylogenetic analysis, consisting of more than 36 TiLV genomes from both newly sequenced and publicly available TiLV genomes, provides new insights into the high genetic diversity of TiLV. This work is an essential steppingstone towards integrating rapid and real-time Nanopore-based amplicon sequencing into routine genomic surveillance of TiLV, as well as future vaccine development.

Nanopore MinION Sequencing Generates a White Spot Syndrome Virus Genome from a Pooled Cloacal Swab Sample of Domestic Chickens in South Africa.

White spot syndrome virus is a highly contagious pathogen affecting shrimp farming worldwide. The host range of this virus is primarily limited to crustaceans, such as shrimps, crabs, prawns, crayfish, and lobsters; however, several species of non-crustaceans, including aquatic insects, piscivorous birds, and molluscs may serve as the vectors for ecological dissemination. The present study was aimed at studying the faecal virome of domestic chickens (Gallus gallus domesticus) in Makhanda, Eastern Cape, South Africa. The cloacal swab specimens (n = 35) were collected from domestic chickens in December 2022. The cloacal swab specimens were pooled-each pool containing five cloacal swabs-for metagenomic analysis using a sequence-independent single-primer amplification protocol, followed by Nanopore MinION sequencing. While the metagenomic sequencing generated several contigs aligning with reference genomes of animal viruses, one striking observation was the presence of a White spot syndrome virus genome in one pool of cloacal swab specimens. The generated White spot syndrome virus genome was 273,795 bp in size with 88.5% genome coverage and shared 99.94% nucleotide sequence identity with a reference genome reported in China during 2018 (GenBank accession: NC_003225.3). The Neighbour-Joining tree grouped South African White spot syndrome virus genome with other White spot syndrome virus genomes reported from South East Asia. To our knowledge, this is the first report of a White spot syndrome virus genome generated from domestic chickens. The significance of White spot syndrome virus infection in domestic chickens is yet to be determined.

Genotypic and antimicrobial susceptibility of Streptococcus agalactiae causing bovine mastitis in the central region of Thailand.

Streptococcus agalactiae is a highly contagious pathogen that causes bovine mastitis, leading to significant economic losses. This study aimed to (1) identify and characterize S. agalactiae strains responsible for bovine mastitis by examining their phenotypic and genotypic characteristics in Thai dairy-intensive farming areas and (2) determine their susceptibility profiles to antimicrobial agents. In total, 100 S. agalactiae isolates obtained from clinical and subclinical mastitis cases from 13 dairy herds located in the central region of Thailand were examined. To confirm the identity of the bacterial pathogens, conventional microbiological procedures recommended by the National Mastitis Council (NMC) and the VITEK® 2 system were employed. All 100 isolates were successfully identified as S. agalactiae using the NMC procedure, whereas 94 isolates were identified as S. agalactiae using the VITEK® 2 system. Finally, the S. agalactiae-specific gene dlt S was identified in all the examined isolates using polymerase chain reaction. Capsular polysaccharide (CPS) typing revealed that all strains belonged to CPS type Ia. Multilocus sequence typing identified 33 selected isolates as sequence type 103. Random amplified polymorphic DNA (RAPD) typing yielded 43 RAPD types, with 6 RAPD clusters identified. These results demonstrated a high level of genetic diversity among S. agalactiae within the studied herds. RAPD analysis suggested that specific S. agalactiae strains could persist in dairy farms for 2-12 months. Furthermore, antimicrobial susceptibility testing was performed using the broth microdilution method. Most strains demonstrated susceptibility to ampicillin, penicillin, penicillin/novobiocin, cephalothin, oxacillin, ceftiofur, and erythromycin. This study revealed the phenotypic and genotypic characteristics of S. agalactiae isolates responsible for bovine mastitis in the central region of Thailand. The rapid identification of S. agalactiae and application of molecular typing methods can provide valuable epidemiological information regarding S. agalactiae causing mastitis in dairy farms. The antimicrobial susceptibility of S. agalactiae indicates that antimicrobial treatment for control and eradication could be a successful protocol. Our findings revealed that a single clonal strain of S. agalactiae affected the 13 studied farms. Further research is needed to explore the feasibility of vaccine development and application.

Alterations of Plasma Biochemical and Immunological Parameters and Spatiotemporal Expression of TLR2 and TLR9 in Gibel Carp (Carassius auratus gibelio) after CyHV-2 Infection.

Cyprinid herpesvirus II (CyHV-2), a highly contagious pathogen of gibel carp (Carassius auratus gibelio), causes herpesviral hematopoietic necrosis disease (HVHND) and enormous financial losses. However, there is limited information available regarding the changes in plasma biochemical and immunological parameters and the response characteristics of Toll-like receptor 2 (TLR2) and Toll-like receptor 9 (TLR9) in gibel carp after CyHV-2 infection. To address this knowledge gap, a sub-lethal CyHV-2 infection was conducted in gibel carp, and the sample was collected daily from 1 to 7 days post infection. The plasma biochemical analyses showed significant decreases in the content of glucose, total cholesterol (TCHO), and total protein (TP), along with marked increases in the level of uric acid, urea, creatinine (CREA), Complement 3 (C3), immunoglobulin D (IgD), and immunoglobulin M (IgM) as well as in the activity of alanine aminotransferase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) in the infected group. Compared with the control group, the concentration of cortisol, triglyceride (TG), and Complement 4 (C4) had no noticeable alterations in the infected group. Real-time quantitative PCR analysis showed significant upregulation of TLR2 and TLR9 mRNA expression in the spleen, kidney, brain, liver, intestine, and gill post CyHV-2 infection. Interestingly, a time- and tissue-dependent expression profile has been comparatively observed for TLR2 and TLR9 in the above tissues of gibel carp after CyHV-2 infection, suggesting distinct roles between TLR2 and TLR9 in antiviral response to CyHV-2 infection. Overall, our results demonstrated that CyHV-2 infection led to the disruption of the physiological metabolic process and damage to the liver and kidney, and induced different spatiotemporal expression patterns of TLR2 and TLR9, ultimately stimulating antiviral response via innate and adaptive immune system. These findings may provide a deeper understanding of the host immunity response to CyHV-2 infection and offer novel perspectives for the prevention and treatment and therapeutic drug development against CyHV-2.

Duck hepatitis A virus type 1 infection induces hepatic metabolite and gut microbiota changes in ducklings

Duck hepatitis A virus type 1 (DHAV-1) can cause severe liver damage in infected ducklings and is a fatal and contagious pathogen that endangers the Chinese duck industry. The objective of this study was to explore the correlation mechanism of liver metabolism-gut microbiota in DHAV-1 infection. Briefly, liquid chromatography-mass spectrometry and 16S rDNA sequencing combined with multivariate statistical analysis were used to evaluate the effects of DHAV-1 infection on liver metabolism, gut microbiota regulation, and other potential mechanisms in ducklings. In DHAV-1-infected ducklings at 72 h postinfection, changes were found in metabolites associated with key metabolic pathways such as lipid metabolism, sugar metabolism, and nucleotide metabolism, which participated in signaling networks and ultimately affecting the function of the liver. The abundance and composition of gut microbiota were also changed, and gut microbiota is significantly involved in lipid metabolism in the liver. The evident correlation between gut microbiota and liver metabolites indicates that DHAV-host gut microbiome interactions play important roles in the development of duck viral hepatitis (DVH).

A novel approach to design a multiepitope peptide as a vaccine candidate for Bordetella pertussis

Bordetella pertussis is a very contagious pathogen in humans, causing pertussis disease. Pertussis is one of the 10 leading causes of death due to infectious diseases, especially among infants and children. Antibiotic-resistant strains have recently emerged in this bacterium, and despite the high vaccination coverage, the prevalence of this disease has been increasing recently in both developed and developing countries. The objective of this study is to introduce a novel in silico vaccine candidate aimed at countering B. pertussis effectively. Differing from other comparable studies, this research employed a computational screening methodology to assess the genome of ‘Bordetella pertussis 18323.’ The purpose was to identify an innovative antigen for the development of a vaccine against B. pertussis. Notably, our investigation introduces an innovative antigen distinguished by its elevated immunogenicity score. Importantly, this antigen lacks toxicity and allergenicity, making it recognizable to the immune system and thus capable of inducing a robust immune response. In the subsequent phase, our antigen was utilized to identify potential epitopes conducive to the construction of a B. pertussis vaccine. These epitopes, alongside linkers, his-tag and adjuvants, were amalgamated to form the vaccine candidate. Subsequently, a comprehensive evaluation of the vaccine was conducted, encompassing various computational tests such as secondary and tertiary structure analysis, physicochemical examination, and structural analysis involving docking and molecular dynamics simulations. Importantly, our vaccine successfully passed all in silico tests. Communicated by Ramaswamy H. Sarma

Insights into varicella-zoster virus assembly from the B- and C-capsid at near-atomic resolution structures

Varicella-zoster is a highly communicable virus that can be transmitted through the airborne route. About one quarter of people are infected with this virus. Previous studies have described the structure of A-capsid and a blurred reconstruction of the C-capsid with icosahedral symmetry. In this study, we have determined the more precise detailed structures of the varicella-zoster virus (VZV) B- and C-capsid in icosahedral symmetry using a combination of block-based reconstruction and symmetry relaxation strategies. In addition, we are reporting structural details of the portal vertex reconstructions in five-fold symmetry and portal reconstructions in twelve-fold symmetry. The structures unveil the basis for the high thermal stability of the VZV capsid. The conformational flexibility of structural elements of the capsid plays a role in the assembly of the capsid and drives processes critical for the viral life cycle. The results of the study open up new avenues for the development of drugs against a highly prevalent and contagious pathogen.

Management Practices of Bovine Mastitis and Milk Quality on Egyptian Dairies.

Milk production continues to be the main source of income for dairy producers, and mastitis continues to be the major health challenge for dairy cows worldwide. The objective of the current study was to describe the different management practices implemented in Egyptian dairies, which may influence mastitis and improve milk quality. An in-person survey was completed with herd managers and owners of 20 Egyptian dairies selected using a stratified random sample from four of Egypt's milk sheds. The questionnaire included 80 questions that inquired about herd demographics, mastitis status and control, milking practices, and management practices of the study dairies. Statistical analyses included descriptive statistics, multiple factor analysis (MFA), and hierarchical clustering to identify the important principal components and different dairy clusters. Of the surveyed herds, 69.50% had less than 500 lactating cows, with Holstein as the main cow breed. The reported average milk production on the study herds was 31.1 (SE ± 1.10) Kg/cow/day. Housing of lactating, dry, and close-up cows was in open lots. The majority of the dairies milked cows were three times per day (90.63%), the remaining milked cows four (5.00%) or a mix of four and three times per day. Furthermore, herringbone parlor design was the most common parlor design (66.79%) in the study dairies. The most common disinfectants used for both pre- and post-milking teat dip were iodine-based, 90.0% and 95.0%, respectively, while 52.16% of the dairies reported that their milkers wore gloves during milking. The reported mean annual percentage of mastitis was 52.3%, as a percent of the milking herd. The study dairies reported the inspection of udder health through either visual inspection (95.00%), udder hygiene score (86.88%), teat end score (71.88%), and/or using CMT (54.91%). Contagious mastitis pathogens were reported in 45.2% of the study herds. More than 50% of the study herds relied on importing pregnant cows or pregnant heifers as replacements. Multiple factor analysis identified 20 questions and represented 5 components of variability related to mastitis on dairies. The current survey of Egyptian dairies described the herd demographics and different management practices related to mastitis control and prevention. All the study dairies relied on the blanket intramammary antimicrobial drugs at dry off when current research elsewhere has identified selective dry cow therapy as an integral component of antimicrobial stewardship on dairies. Further research is required to identify the association between different management factors and the occurrence of mastitis.

A novel live attenuated vaccine candidate protects chickens against subtype B avian metapneumovirus

Avian metapneumovirus (aMPV) is a highly contagious pathogen that causes acute upper respiratory tract diseases in chickens and turkeys, resulting in serious economic losses. Subtype B aMPV has recently become the dominant epidemic strain in China. We developed an attenuated aMPV subtype B strain by serial passaging in Vero cells and evaluated its safety and efficacy as a vaccine candidate. The safety test showed that after the 30th passage, the LN16-A strain was fully attenuated, as clinical signs of infection and histological lesions were absent after inoculation. The LN16-A strain did not revert to a virulent strain after five serial passages in chickens. The genomic sequence of LN16-A differed from that of the parent wild-type LN16 (wtLN16) strain and had nine amino acid mutations. In chickens, a single immunization with LN16-A induced robust humoral and cellular immune responses, including the abundant production of neutralizing antibodies, CD4+ T lymphocytes, and the Th1 (IFN-γ) and Th2 (IL-4 and IL-6) cytokines. We also confirmed that LN16-A provided 100% protection against subtype B aMPV and significantly reduced viral shedding and turbinate inflammation. Our findings suggest that the LN16-A strain is a promising live attenuated vaccine candidate that can prevent infection with subtype B aMPV.

The Detection of Bacterial Pathogens, including Emerging Klebsiella pneumoniae, Associated with Mastitis in the Milk of Ruminant Species

Milk is an excellent medium for the growth of several bacteria and other microorganisms and thus, it has been extensively studied. An always current issue in the dairy industry is mastitis, which causes losses in milk volume and profits. In many cases, milk is used raw or treated at low temperatures for further dairy processes while there are quite a few cases in which foodborne-related outbreaks have occurred. Both culture-based methods and PCR were used to assess the presence of certain pathogens related to both contagious and environmental pathogens, especially the emerging pathogenic bacterium Klebsiella pneumoniae, as well as Staphyloccocus aureus and Enterobacter spp., which are associated with mastitis in milk samples from different lactating ruminant species (cows, goats, and sheep) and to further evaluate the significance of the isolated pathogens to public health. Even though significant mastitis contagious pathogens such as Staphylococcus aureus and Staphylococcus epidermidis were not detected, environmental pathogens related to poor hygiene conditions at the farm level (K. pneumoniae, Staphylococcus saprophyticus, and Enterococcus spp.) were detected. In particular, K. pneumoniae and Staphylococcus saprophyticus were present in ovine milk samples while bovine and caprine milk samples were contaminated with Enterococcus spp. The presence of these bacteria underlines the significant role of environmental hygiene especially since Staphylococcus saprophyticus and Enterococcus spp. are related to urinary tract infections and all of the tested pathogens may carry antibiotic resistance genes. More specifically, 20% of the isolated Klebsiella pneumoniae strains were found resistant to carbapenem antibiotics. The presence of emerging K. pneumoniae in ovine milk samples also indicates the need for new policies in terms of safety testing. Suggestions of monitoring processes carried out by the relevant authorities are discussed.

The US Biological and Chemical Agression Against Cuba in 1963–1996

For almost 60 years, the United States has tried to overthrow the government of Fidel Castro in Cuba, using a variety of means, including chemical and biological agents. At the same time, their use has always been officially denied by Washington. The purpose of this article is to summarize the available sources proving the involvement of the United States, as well as the groups of the Cuban opposition created and supported by the US, in biological and chemical sabotage against both the economy and the people of Cuba. Sources, research methods. The author used a variety of open sources, including the statements by Cuban officials, materials from the Cuban and American press, declassified documents from the US government agencies, as well as public confessions of former CIA agents and Cuban opposition figures. The research method is the cross analysis. The discussion of the results. The declassified US documents show that the organizers of «Operation 40» and «Operation Mongoose» planned to use biological agents and parasitic plants against plantations of crops and livestock. Sugar exported to the USSR was also poisoned. In addition, the head of one of the Cuban terrorist organizations admitted that he had imported biological agents to Cuba, which provoked an epidemic of dengue hemorrhagic fever in 1981. The testimoies of several other former CIA agents also point to the use of African swine fever in 1971 to kill pigs in Cuba. The atypical course of the Dengue fever epidemic in 1981, as well as the absence of a strain (Dengue-2) of this virus in the Western Hemisphere at that time, indicate the planned nature of the sabotage, which resulted in an outbreak on the island. Conclusions. During the Cold War, the United States actively used chemical and biological agents against agriculture and the people of Cuba. The facts considered allow us to draw conclusions about the nature of the methods that the CIA used to organize and conduct biosabotage: such operations were planned «in the long run» and had large-scale goals – for example, to eliminate sugar production on the island or cause famine; highly contagious pathogens of human and animal diseases were used for biological sabotage; artificially caused outbreaks of epidemics and epizootics were disguised as natural ones; representatives of the local opposition, as well as mercenaries, were involved in the implementation of sabotage; to infect farm animals, vaccines contaminated with virulent strains directly at biofactories, as well as contaminated feed were used.

Quantitative analysis of acetylation in peste des petits ruminants virus-infected Vero cells

BackgroundPeste des petits ruminants virus (PPRV) is a highly contagious pathogen that strongly influences the productivity of small ruminants worldwide. Acetylation is an important post-translational modification involved in regulation of multiple biological functions. However, the extent and function of acetylation in host cells during PPRV infection remains unknown.MethodsDimethylation-labeling-based quantitative proteomic analysis of the acetylome of PPRV-infected Vero cells was performed.ResultsIn total, 1068 proteins with 2641 modification sites were detected in response to PPRV infection, of which 304 differentially acetylated proteins (DAcPs) with 410 acetylated sites were identified (fold change < 0.83 or > 1.2 and P < 0.05), including 109 up-regulated and 195 down-regulated proteins. Gene Ontology (GO) classification indicated that DAcPs were mostly located in the cytoplasm (43%) and participated in cellular and metabolic processes related to binding and catalytic activity. Functional enrichment indicated that the DAcPs were involved in the minichromosome maintenance complex, unfolded protein binding, helicase activity. Only protein processing in endoplasmic reticulum pathway was enriched. A protein-protein interaction (PPI) network of the identified proteins further indicated that a various chaperone and ribosome processes were modulated by acetylation.ConclusionsTo the best of our knowledge, this is the first study on acetylome in PPRV-infected host cell. Our findings establish an important baseline for future study on the roles of acetylation in the host response to PPRV replication and provide novel insights for understanding the molecular pathological mechanism of PPRV infection.

Brucellae as resilient intracellular pathogens: epidemiology, host-pathogen interaction, recent genomics and proteomics approaches, and future perspectives.

Brucellosis is considered one of the most hazardous zoonotic diseases all over the world. It causes formidable economic losses in developed and developing countries. Despite the significant attempts to get rid of Brucella pathogens in many parts of the world, the disease continues to spread widely. Recently, many attempts proved to be effective for the prevention and control of highly contagious bovine brucellosis, which could be followed by others to achieve a prosperous future without rampant Brucella pathogens. In this study, the updated view for worldwide Brucella distribution, possible predisposing factors for emerging Brucella pathogens, immune response and different types of Brucella vaccines, genomics and proteomics approaches incorporated recently in the field of brucellosis, and future perspectives for prevention and control of bovine brucellosis have been discussed comprehensively. So, the current study will be used as a guide for researchers in planning their future work, which will pave the way for a new world without these highly contagious pathogens that have been infecting and threatening the health of humans and terrestrial animals.

Pantoea agglomerans: A rare infectious outbreak affecting maintenance hemodialysis patients in a tertiary care hospital.

Pantoea agglomerans is an environmental pathogen known to cause infection in immunocompromised individuals, particularly after thorn injuries. However, previous data showed few cases of human disease caused by contaminated medical products such as parenteral nutrition, anesthetic agents, blood, and peritoneal dialysis solutions. Infection in hemodialysis patients is rare. In this study, we presented a detailed account of several hemodialysis patients infected with this contagious pathogen and compared them with noninfected dialysis patients. We retrospectively reviewed the hospital records of 105 hemodialysis patients. Seventeen of 105 patients were diagnosed with P. agglomerans infection. We carefully analyzed their entire in-hospital course. Among infected patients, 52.9% were male with a median age of 49 (IQR: 32-66) years. Compared to the noninfected patients, age below 50 years, prior kidney transplantation, prior immunosuppression and antibiotics use, and dialysis via a tunneled vascular catheter were the significant epidemiological features. Despite negative microbiological investigations, we suspect the possible infectious spread via infected central venous catheter was the likely infectious source. Most importantly, all patients responded well to intravenous antibiotics. Only two patients required the removal of the tunneled catheter. Their mortality rate was 0%. P. agglomerans infection, although considered rare, is becoming increasingly prevalent among dialysis patients. Its occurrence must be appraised as an infectious outbreak rather than mere contamination. Prompt treatment, source identification, and early implementation of preventive strategies should always be the goal to curtail this infection at an early stage.

Virtual Screening, ADMET Analysis, and Synthesis of 2-(1H-benzotriazol- 1-yl) N- substituted Acetohydrazide that Binds to the Glycoprotein B of Herpes Simplex Virus-I (HSV-I)

Background: Herpes simplex Virus type 1 (HSV-1) is a contagious human pathogen causing severe infection. In recent decades, the virus has become dormant and resistant to available treatment creating the need for the development of new therapeutic agents against it. Benzotriazole is a versatile molecule with a wide range of activities like antibacterial, antiprotozoal, antiulcer, anthelmintic, and antiproliferative activity. Methods: A series of 2-(1H- benzotriazole-1-yl) N- substituted acetohydrazide derivatives were synthesized using the method given in the literature. The derivatives were obtained in good yield and characterized by spectral methods of analysis. The antiviral activity against the glycoprotein B of Herpes Simplex Virus-I (HSV-I) was determined using molecular docking (2GUM). Result: All compounds had strong binding affinity over the standard Acyclovir. Compound 5h had the highest binding affinity and the highest inhibitory activity. Conclusion: The Benzotriazole-N- substituted acetohydrazide derivatives has the highest bind-ing affinity and good inhibition of glycoprotein B of Herpes Simplex Virus-I (HSV-I), which makes it a good antiviral agent.

Understanding rabbit owners' willingness to engage in disease prevention behaviors.

Rabbit hemorrhagic disease virus 2 (RHDV2) is a fatal, highly contagious pathogen that infects wild and domestic lagomorphs (rabbits and hares). RHDV2 is an important cause of disease in pet and companion rabbits, has resulted in economic losses for the commercial rabbit industry, and has caused declines of wild lagomorph populations. It is essential for domestic rabbit owners to engage in appropriate actions (e.g., using effective disinfectants, creating secure barriers between domestic and wild rabbits) to protect the health and welfare of their rabbits and reduce the risk of human-mediated spread of RHDV2. Thus, we investigated rabbit owners' stated willingness to engage in nine commonly recommended biosecurity practices and their support for seven potential government-implemented management actions. We administered an online survey to 1790 rabbit owners in the United States between April and August 2021. Respondents were likely to engage in all biosecurity measures and were supportive of most management actions that could be implemented by government agencies. Respondents' willingness to engage in and support biosecurity measures was positively correlated with their perceptions of the importance of biosecurity, risk perceptions pertaining to the impact of RHDV2 on lagomorphs and rabbit-related industries, knowledge of RHDV2, and trust in government to manage RHDV2. Respondents' motivations for owning rabbits, husbandry behaviors, and demographic characteristics also influenced their willingness to engage in or support biosecurity measures. Engaging domestic rabbit owners in collaborative biosecurity measures is critical for protecting domestic rabbit health and preventing potential spillover between domestic and free-roaming lagomorphs, as there are still many uncertainties about how RHDV2 is spreading across the United States and the world. Implementing outreach strategies that communicate the importance and effectiveness of biosecurity practices in protecting rabbit welfare, rabbit-related activities, and wild lagomorph populations may increase the likelihood of rabbit owners adopting biosecurity measures.

Combined regulation of pro-inflammatory cytokines production by STAT3 and STAT5 in a model of B. pertussis infection of alveolar macrophages.

Bordetella pertussis is a highly contagious respiratory pathogen responsible for whooping-cough or pertussis. Despite high vaccination coverage worldwide, this gram-negative bacterium continues to spread among the population. B. pertussis is transmitted by aerosol droplets from an infected individual to a new host and will colonize its upper respiratory tract. Alveolar macrophages (AMs) are effector cells of the innate immune system that phagocytose B. pertussis and secrete both pro-inflammatory and antimicrobial mediators in the lungs. However, understanding their role in B. pertussis pathogenesis at the molecular level is hampered by the limited number of primary AMs that can be collected in vivo. In order to decipher the regulation of innate response induced by B. pertussis infection, we used for the first time self-renewing, non-transformed cells, called Max Planck Institute (MPI) cells, which are phenotypically and functionally very close to pulmonary AMs. Using optimized infection conditions, we characterized the entry and the clearance of B. pertussis within MPI macrophages. We showed that under these conditions, MPI cells exhibit a pro-inflammatory phenotype with the production of TNF, IL-1β, IL-6 and MIP-2α, similarly to primary AMs purified from broncho-alveolar fluids of mice. In addition, we explored the yet uncharacterized role of the signal transduction activator of transcription (STAT) proteins family in the innate immune response to B. pertussis infection and showed for the first time the parallel regulation of pro-inflammatory cytokines by STAT3 and STAT5 in MPI macrophages infected by B. pertussis. Altogether, this work highlights the interest of using MPI cells for experiments optimization and preliminary data acquisition to understand B. pertussis interaction with AMs, and thus significantly reduce the number of animals to be sacrificed.