Animal Pathogens Research Articles

Leveraging Lactobacillus plantarum probiotics to mitigate diarrhea and Salmonella infections in broiler chickens

Anaerobic bacteria, such as Lactobacillus plantarum (LP), are known to play a significant role in maintaining gut health and protecting against enteric pathogens in animals. The present study aimed to develop a safe, affordable, and eco-friendly method for producing LP-based probiotics and evaluate their efficacy in mitigating Salmonella-induced diarrhea in broiler chickens. The study employed three different culture media (MRS, TSB, and Baird Parker) to grow LP, which was then dried using a spray-drying technique to produce a stable probiotic formulation. When administered to broiler chickens, the LP probiotic derived from the MRS medium significantly improved body weight gain (4.147-fold increase over 4 weeks) compared to the other two culture conditions. Importantly, the LP probiotic treatment could substantially reduce the diarrhea index in broilers, with up to an 86.45% improvement in Salmonella-induced enteric infections. The beneficial effects were attributed to the ability of LP to modulate the gut microbiome, enhance the integrity of the intestinal mucosa, and mitigate the pathogenic effects of Salmonella. These findings demonstrate the potential of anaerobic Lactobacillus plantarum as a safe and effective probiotic intervention for controlling enteric diseases and improving production outcomes in poultry farming. The developed method provides a sustainable approach to harness the beneficial properties of this anaerobic bacterium for animal health and welfare.Graphical abstract

Aggregation-Induced Emission Luminogens for Plant Photodynamic Seed Sterilization.

Pathogen-carrying seeds can significantly impact plant growth and development and may lead to serious public health incidents. Modern agriculture heavily relies on synthetic chemical microbicides and physical methods to eradicate pathogens transmitted by plant seeds. To counteract the misuse of microbicides, a class of cationic amphiphilic aggregate-induced emission luminogens (AIEgens) are developed as photodynamic seed sterilization agents. AIEgens function as antimicrobial agents in seed treatment. These materials are engineered to specifically bind to pathogenic microorganisms on seed surfaces. Furthermore, when combined with photodynamic therapy, AIEgens can be activated to produce reactive oxygen species that selectively destroy pathogens. Sterilization experiments with tomato seeds carrying Pseudomonas syringae and mung bean seeds carrying Pseudomonas aeruginosa demonstrate that AIEgens can effectively eliminate both plant and animal pathogens carried by seeds. Therefore, AIEgens offer a promising solution for preventing the spread of seed-borne pathogens.

Structural characterization of the full-length Hantaan virus polymerase.

Hantaviridae are a family of segmented negative-sense RNA viruses that contains important human and animal pathogens. Hantaviridae contain a viral RNA-dependent RNA polymerase that replicates and transcribes the viral genome. Here we establish the expression and purification polymerase from the Old World Hantaan virus and characterise the structure using Cryo-EM. We determine a series of structures at resolutions between 2.7 and 3.3 Å of RNA free polymerase comprising the core, core and endonuclease, and a full-length polymerase. The full-length polymerase structure depicts the location of the cap binding and C-terminal domains which are arranged in a conformation that is incompatible with transcription and in a novel conformation not observed in previous conformations of cap-snatching viral polymerases. We further describe structures with 5' vRNA promoter in the presence and absence of a nucleotide triphosphate. The nucleotide bound structure mimics a replication pre-initiation complex and the nucleotide stabilises the motif E in a conformation distinct from those previously observed. We observe motif E in four distinct conformations including β-sheet, two helical arrangements, and nucleotide primed arrangement. The insights gained here guide future mechanistic studies of both the transcription and replication activities of the hantavirus polymerase and for the development of therapeutic targets.

ISOLATION, BIOCHEMICAL CHARACTERIZATION AND ANTIBIOTIC SENSITIVITY OF LACTOBACILLI IN QUAIL DROPPINGS

Background: Probiotics are microorganisms which are when given in a specific amount, have favorable effects on the host health. Objectives: The current study was conducted on quail’s droppings, which were collected from the various sailing points within district Dera Ismail Khan. The core aim of this field study was to isolate the Lactobacillus species from the droppings to identify and characterize them. Methods: Three species (lactobacillus brevis, lactobacillus salivarius and lactobacillus fermentum) were isolated on the basis of biochemical tests. Results: Lactobacillus salivarius was found more tolerant at pH 2 as compared to the other two species of lactobacillus, likewise at pH 3 lactobacillus brevis showed maximum growth, which was significantly higher than other species of lactobacillus. Similarly, all the three species of lactobacillus showed higher growth on 0.5 % ox gall as compared to the 1 % ox gall. Likewise antibiotic sensitivity testing of lactobacillus species shown sensitivity to erythromycin except L. fermentum and all three were resistance to tetracycline, kanamycin, ciprofloxacin and gentamycin. Conclusion: It is concluded that lactobacillus species have huge benefits like their tolerance to low pH and their growth on bile salts. Their use in diets will not only balance the gut microflora but also will inhibit the growth of pathogens in humans, animals and poultry species, as described by different studies which are conducted on above mentioned species.

High Occurrence of ST131 Among Extraintestinal Pathogenic Escherichia coli in Humans and Dogs in Sri Lanka.

Extraintestinal pathogenic Escherichia coli (ExPEC) is an important human and animal pathogen. In particular, the ST131 of ExPEC is a widely distributed clone, resistant to extended-spectrum cephalosporins and fluoroquinolones. We investigated the occurrence of ST131 among ExPEC from humans and dogs in Sri Lanka and determined its antimicrobial resistance. A total of 215 ExPEC isolates were collected from humans (n = 179) and dogs (n = 36) from two different cities in Sri Lanka. Antimicrobial resistance was tested by the disk diffusion method, and the presence of resistance-encoding genes (blaTEM, blaSHV and blaCTX-M) and ST131 clades/subclades was tested by PCR. The majority of the isolates were (61.8%) ST131, with 55.8% in ST131-clade C. Of the clade C isolates, 44.3%, 27.8%, 15.7% and 6.7% were in clades C2, C other, C1-non-M27 and C1-M27, respectively. Approximately 11% of the ST131 isolates were carbapenem resistant. ExPEC from dogs showed comparable resistance rates to human isolates except for resistance to amoxicillin-clavulanate and amikacin. We observed a high occurrence of ST131 and its clade C, with more than 11% exhibiting resistance to carbapenems in Sri Lanka. Furthermore, ST131-C1-M27, with high resistance to both quinolones and extended-spectrum cephalosporins, was also present. Our results emphasise the importance of the One Health approach in the monitoring of antimicrobial-resistant E. coli in different regions of Sri Lanka to gain a better understanding of their prevalence over time, contributing to effective antimicrobial stewardship.

Development and validation of animal variant classification guidelines to objectively evaluate genetic variant pathogenicity in domestic animals

Assessing the pathogenicity of a disease-associated genetic variant in animals accurately is vital, both on a population and individual scale. At the population level, breeding decisions based on invalid DNA tests can lead to the incorrect inclusion or exclusion of animals and compromise the long-term health of a population, and at the level of the individual animal, lead to incorrect treatment and even life-ending decisions. Criteria to determine pathogenicity are not standardized, i.e., no guidelines for animal variants are available. Here, we aimed to develop and validate guidelines to be used by the community for Mendelian disorders in domestic animals to classify variants in categories based on standardized criteria. These so-called animal variant classification guidelines (AVCG) were based on those developed for humans by The American College of Medical Genetics and Genomics (ACMG). In a direct comparison, 83% of the pathogenic variants were correctly classified with ACMG, while this increased to 92% with AVCG. We described methods to develop datasets for benchmarking the criteria and identified the most optimal in silico variant effect predictor tools. As the reproducibility was high, we classified 72 known disease-associated variants in cats and 40 other disease-associated variants in eight additional species.

Exploring the Gastrointestinal Microbiome of Eurasian Griffon Vultures (Gyps fulvus) Under Rehabilitation in Portugal and Their Potential Role as Reservoirs of Human and Animal Pathogens

The Eurasian griffon vulture (Gyps fulvus), a widely distributed scavenger, plays a crucial role in ecosystem health by consuming decomposing carcasses. Scavengers have adapted to avoid disease from the rotting carrion they feed on, probably through a specialized gut microbiome. This study aimed to characterize the gut microbiome of G. fulvus (n = 8) present in two rehabilitation centers in mainland Portugal and evaluate their potential as reservoirs of pathogens. Samples were studied through high-throughput 16S rDNA amplicon sequencing of the hypervariable V3-V4 regions and further analyzed using the Qiime2 bioinformatics platform. Our results showed that factors such as sex, location, and time of sampling did not significantly affect the gut microbiome of the griffon vulture. Its composition was highly similar to that of phylogenetically closed animals. However, several potential human and veterinary pathogens were identified. In conclusion, the gut microbiome of Gyps fulvus in rehabilitation centers is not significantly altered by stress associated with captivity. Its composition is similar to that of other vultures and scavengers due to their identic diet and needs, suggesting a well-conserved functional gut microbiome, which seems to be influenced by season. The potential risks posed by the identified pathogens to humans and other animals should be further investigated.

Genetic diversity of populations of Simulium reptans s.l. (Diptera: Simuliidae) in the territory of Russia and Northern Kazakhstan.

The presence of sibling (cryptic) species among bloodsucking black flies can be a serious problem in assessing both their diversity and the role of individual species in the transmission of dangerous human and animal pathogens. The peculiarities of molecular genetic structure of populations and ecology of black flies of Simulium reptans s.l. (complex of Simulium reptans (Linnaeus) and Simulium reptantoides Carlsson) in European and Asian parts of Russia and Northern Kazakhstan in comparison with populations from Central and Northern Europe were investigated. Using restriction analysis of the COI gene and subsequent sequencing, we searched for unique mtDNA variants that could belong to previously undescribed forms of the studied group. In the north of the European part of Russia (north of Eastern Europe), only S. reptans A was registered, and in the Asian part of Russia (Siberia) and Northern Kazakhstan, only S. reptans B was found. Using phylogenetic analysis of the total data, three branches of S. reptans B were revealed and named in accordance with the areas of their distribution-East European, Eurasian and Asian. Simulium reptantoides Carlsson has currently been found only in Europe. The peculiarities of distribution and possible adaptations to different habitats of S. reptantoides and representatives of individual groups within S. reptans are discussed. Based on the characteristics of the sampling sites of the studied specimens, as well as the samples available in the NCBI database, the ecological and geographical features of the habitats of the molecular forms of Simulium reptans A and B were revealed. To obtain more accurate information on the distribution of S. reptans A and B forms in Eurasia, taking into account the elevational zonation, it is necessary to conduct additional studies in the mountainous regions of South-Western Europe, Siberia and Northern China. Based on the sequences presented in the DNA database, a method (AS-PCR COI variant) for separating the cryptic species-Simulium reptans (Linnaeus) and S. reptantoides Carlsson-was proposed. During testing of this method on the material collected in Russia and Northern Kazakhstan, only S. reptans was identified (S. reptantoides was absent), which completely coincided with the sequencing results. To further confirm the effectiveness of the method, it is necessary to conduct testing on material including representatives of the both species.

Parasitic Protozoa and Other Vector-Borne Pathogens in Captive Mammals from Brazil

In captive environments, mammals are frequently exposed to various parasitic protozoa and other vector-borne pathogens that can impact both animal health and public health. Monitoring these pathogens is essential for animal welfare and zoonotic disease control. This study aimed to investigate the prevalence of parasitic protozoa and other vector-borne pathogens in captive mammals through molecular detection methods at the Belo Horizonte Zoo, Brazil. Between November 2021 and March 2023, whole blood samples were collected from 40 mammals. Molecular analyses identified piroplasms, Leishmania spp., granulocytic/platelet Anaplasma/Ehrlichia spp., monocytic Ehrlichia spp., Bartonella spp. and hemotropic Mycoplasma spp. with a 72.5% positivity rate. Piroplasms were found in 22.5% (two Pantanal cats, two gorillas, one white rhinoceros, one spider monkey, one jaguar, one tufted capuchin and one hippo) and Leishmania spp. in 12.9% (four maned wolves). Granulocytic/platelet Anaplasma/Ehrlichia spp. were found in 12.5% of the samples (one gorilla and four maned wolves), Ehrlichia canis in 2.5% of the animals (one maned wolf), Bartonella spp. in 42.5% (six howler monkeys, two maned wolves, one gorilla, one white rhino, one southern tamandua, one common woolly monkey, one tufted capuchin, one brown brocket deer, one agouti, one cougar and one hippo), hemotropic Mycoplasma spp. in 17.5% (one gorilla, one maned wolf, one white rhino, one howler monkey, two common woolly monkeys and one European fallow deer). Five Artiodactyla members tested negative for A. marginale. Coinfections occurred in 34.5% of the positive samples. Sequencing revealed that Theileria spp. and Cytauxzoon spp. are closely related to Theileria bicornis and Cytauxzoon felis; Ehrlichia canis and Bartonella spp. are closely related to B. clarridgeiae and B. henselae; and hemotropic Mycoplasma spp. are closely related to Candidatus Mycoplasma haemominutum. Our results showed a high occurrence of vector-borne pathogens in captive animals, including zoonotic species, which may pose a risk to animal and human public health.

Whole genome sequencing reveals circulation of potentially virulent Listeria innocua strains with novel genomic features in cattle farm environments in Dhaka, Bangladesh.

Through the last decade, Listeria spp. has been detected in food and environmental samples in Bangladesh. However, the genomic information of this bacterium that prevails in the country remains scarce. This study analyzed the complete genome sequences of two Listeria spp. isolates obtained from cow dung and their drinking water collected from a cattle farm in Dhaka, Bangladesh. Both the isolates were identified as Listeria innocua, which shared almost identical genomic features. The genome sequences demonstrated the presence of 13 virulence genes associated with invasion (iap/cwhA, gtcA, and lpeA), surface protein anchoring (lspA), adherence (fbpA, and lap), intracellular survival (lplA1, and prsA2), peptidoglycan modification (oatA, and pdgA), and heat stress (clpC, clpE, and clpP). Additionally, the gene fosX, conferring resistance to fosfomycin, and two copper resistance-associated genes, copC and csoR, were identified in both. The genome sequences also revealed two plasmid replicons, rep25 and rep32, along with three insertion sequences [ISLmo3 (CP022021), ISLmo7 (CP006611), ISS1N (M37395)]. Notably, a composite transposon [CN_8789_ISS1N (M37395)], was detected in both L. innocua isolates, representing the first documented occurrence of this particular composite transposon in any reported Listeria species. Furthermore, the genomes contained four prophage regions [Listeria phage LP-030-2 (NC_021539), Listeria phage vB_LmoS_188 (NC_028871), Listeria phage A118 (NC_003216) and Escherichia phage RCS47 (NC_042128)]. Two CRISPR arrays were also identified, one belonging to the family type II-A. Multilocus Sequence Typing (MLST) analysis classified the L. innocua isolates of the same sequence type, ST-637. Single nucleotide polymorphism (SNP) analysis uncovered the presence of 231-340 SNPs between the L. innocua isolates and their closely related global lineage. In contrast, only 42 SNPs were identified between the two isolates, suggesting a potential transmission of L. innocua between cow dung and cattle farm water. The presence of L. innocua isolates harboring virulence genes associated with ruminant infection in the cattle farm environment of Bangladesh raises significant concerns about the potential presence of other human and animal pathogens. This poses a serious threat to the cattle farming industry. Additionally, the genomic analysis of the L. innocua isolates enhances our understanding of the evolutionary dynamics of Listeria species.

Detection of selected vector-borne pathogens in domestic animals, ectoparasites, and their owners in a rural community in Southwest Guatemala

Detection of selected vector-borne pathogens in domestic animals, ectoparasites, and their owners in a rural community in Southwest Guatemala

Occurrence of Staphylococcus aureus, Staphylococcus epidermidis, and Staphylococcus pseudintermedius colonization among veterinarians in the province of Malaga, Spain

Background and Aim: Staphylococcus pseudintermedius and Staphylococcus aureus are common colonizing pathogens in companion animals. These opportunistic pathogens can cause infections of varying frequency and severity in humans and pets. Studies on Staphylococcus colonization in veterinarians are scarce. This study aimed to investigate the colonization of the nostrils and hands by S. aureus, Staphylococcus epidermidis, and S. pseudintermedius among healthy clinical practice veterinarians in the province of Malaga (Spain), with a particular focus on their potential antibiotic resistance. Materials and Methods: A request for voluntary participation was extended to professionals from the Official College of Veterinarians of Malaga. Nasal and hand swabs were collected by two trained technicians in January 2024, and all samples were delivered to the laboratory within 24 h. Gram staining, catalase, oxidase, and coagulase tests were performed. The susceptibility of the isolated bacteria to 11 antibiotics was evaluated. Results: A total of 50 clinical practice veterinarians were enrolled in the study, comprising 36 women and 14 men from 31 veterinary clinics across Málaga province. A total of 32% of the nasal samples yielded S. aureus, whereas 64% were found to contain S. epidermidis. In total, 30% of the hand samples yielded S. aureus and 30% yielded S. epidermidis. The participants did not exhibit any strains of S. pseudintermedius in their nasal samples or hands. Two strains (11.1%) of methicillin-resistant S. aureus were isolated from 18 strains isolated from nostrils. Furthermore, a high prevalence of S. aureus strains resistant to ampicillin (94.4%) and amoxicillin (72.2%) was observed. Conclusion: The colonization profiles of veterinary professionals were similar to those observed in the general population. Further research is required among veterinary professionals, companion animals, and their owners to better understand the colonization processes and the pet-human interface within a “One Health” approach. Keywords: antibiotic resistance, drug-resistant Staphylococcus aureus, one health, Seroprevalence, Staphylococcus colonization, veterinarians.

Legionella pneumophila IrsA, a novel, iron-regulated exoprotein that facilitates growth in low-iron conditions and modulates biofilm formation.

To discover new factors that are involved in iron acquisition by Legionella pneumophila, we used RNA-Seq to identify the genes that are most highly induced when virulent strain 130b is cultured in a low-iron chemically defined medium. Among other things, this revealed 14915, a heretofore uncharacterized gene that is predicted to be transcriptionally regulated by Fur and to encode a novel, ~15 kDa protein. 14915 was present in all L. pneumophila strains examined and had homologs in a subset of the other Legionella species. Compatible with it containing a classic signal sequence, the 14915 protein was detected in bacterial culture supernatants in a manner dependent upon the L. pneumophila type II secretion system. Thus, we designated 14915 as IrsA for iron-regulated, secreted protein A. Based on mutant analysis, the irsA gene was not required for optimal growth of strain 130b in low-iron media. However, after discovering that the commonly used laboratory-derived strain Lp02 has a much greater requirement for iron, we uncovered a growth-enhancing role for IrsA after examining an Lp02 mutant that lacked both IrsA and the Fe2+-transporter FeoB. The irsA mutant of 130b, but not its complemented derivative, did, however, display increased biofilm formation on both plastic and agar surfaces, and compatible with this, the mutant hyper-aggregated. Thus, IrsA is a novel, iron-regulated exoprotein that modulates biofilm formation and, under some circumstances, promotes growth in low-iron conditions. For this study, we determined and deposited in the database a complete and fully assembled genome sequence for strain 130b.IMPORTANCEThe bacterium Legionella pneumophila is the principal cause of Legionnaires' disease, a potentially fatal form of pneumonia that is increasing in incidence. L. pneumophila exists in many natural and human-made water systems and can be transmitted to humans through inhalation of contaminated water droplets. L. pneumophila flourishes within its habitats by spreading planktonically, assembling into biofilms, and growing in larger host cells. Iron acquisition is a key determinant for L. pneumophila persistence in water and during infection. We previously demonstrated that L. pneumophila assimilates iron both by secreting a non-protein iron chelator (siderophore) and by importing iron through membrane transporters. In this study, we uncovered a novel, secreted protein that is highly iron-regulated, promotes L. pneumophila's growth in low-iron media, and impacts biofilm formation. We also identified uncharacterized, IrsA-related proteins in other important human and animal pathogens. Thus, our results have important implications for understanding iron assimilation, biofilm formation, and pathogenesis.

Importance of benzoyltransferase GcnE and lysine benzoylation of alcohol dehydrogenase AdhB in pathogenesis and aflatoxin production in Aspergillus flavus.

Lysine benzoylation (Kbz) is a newly identified post-translational modification associated with active transcription and metabolism in eukaryotes. However, whether Kbz exists in pathogenic fungi and its function remains unknown. Here, we demonstrated for the first time that Kbz is present in Aspergillus flavus and identified 60 benzoylated sites on 46 benzoylated proteins by global benzoylome analysis. Our data demonstrated that alcohol dehydrogenase B (AdhB) is regulated by benzoylation on lysine 321 (K321), and mutations of Kbz site in AdhB significantly reduced the alcohol dehydrogenase activity in vivo and in vitro. Both adhB deletion mutant and benzoylated site mutants (K321R and K321A) exhibited similar phenotype, including decreased conidiation and seed colonization, increased sclerotia formation and aflatoxin production, and more sensitive to cell wall damage stress. We also found that GcnE has benzoyltransferase activity in vitro and in vivo, and its repression leads to decreased Kbz level and enzymatic activity of AdhB. The catalytic site E139 is important for the benzoyltransferase function of GcnE. Our study uncovers a previously unknown mechanism by which benzoylation regulates AdhB activity to affect the development, secondary metabolism, pathogenicity, and stress response of A. flavus. Meanwhile, it points out the important role of Kbz in the pathogenicity of pathogenic fungi.IMPORTANCEAspergillus flavus is a ubiquitous opportunistic pathogen of plants and animals, which produces carcinogenic and toxic secondary metabolite aflatoxin. A. flavus and aflatoxin contamination have emerged as a global food safety concern. Currently, post-translational modification plays crucial modulatory roles in the fungal development and virulence, but the role of benzoylation in fungal pathogenicity remains undetermined, which limits the development of prevention and control technique. Here, we first identified 46 benzoylated proteins in A. flavus, and found that benzoyltransferase GcnE exerted effects on pathogenicity and aflatoxin production by regulating the benzoylation of AdhB. This finding not only provided valuable information for prevention and control of A. flavus contamination, but also offered basic knowledge for investigation of the regulation mechanism of secondary metabolism in other fungi.

Genome-wide quantification of polycistronic transcription in Leishmania major.

Leishmania major is a human-pathogenic, obligate parasite and the etiological agent of the most prevalent, cutaneous form of leishmaniasis, which is an important neglected, tropical disease with ~1.2 million new infections per year. Leishmania, and the whole order Trypanosomatida, are early eukaryotes with highly diverged gene expression and regulation pathways, setting them apart from their mammalian hosts and from most other eukaryotes. Using precision run-on sequence analysis, we performed a genome-wide mapping and density analysis of RNA polymerases in isolated nuclei of the protozoan parasite Leishmania major. We map transcription initiation sites at divergent strand switch regions and head-tail regions within the chromosomes and correlate them with known sites of chromatin modifications. We confirm continuous, polycistronic RNA synthesis in all RNA polymerase II-dependent gene arrays but find small varying RNA polymerase activities in polycistronic transcription units (PTUs), excluding gene-specific transcription regulation, but not PTU-specific variations. Lastly, we find evidence for transcriptional pausing of all three RNA polymerase classes, hinting at a possible mechanism of transcriptional regulation.IMPORTANCELeishmania spp. are pathogens of humans and animals and cause one of the most important neglected tropical diseases. Regulation of gene expression in Leishmania but also in the related Trypanosoma is radically different from all eukaryotic model organisms, dispensing with regulated, gene-specific transcription, and relying instead on highly regulated translation. Our work sheds light on the initiation, elongation, and termination of transcription, maps unidirectional, polycistronic transcription units, provides evidence for transcriptional pausing at or near starting points of RNA synthesis, and quantifies the varying transcription rates of the polycistronic transcription units. Our results will further the understanding of these important pathogens and should provide a valuable resource for researchers in the field of eukaryotic microbiology.

Use of a Novel Feeding System to Assess the Survival of a Very Stable Mammalian Virus, Porcine Parvovirus, Within Black Soldier Fly (Hermetia illucens) Larvae: A Comparison with Mealworm (Tenebrio molitor) Larvae

Insect larvae production offers the potential for large-scale synthesis of high-quality protein that can be used as feed or food. However, currently, there are limitations on the source of substrates for the insect larvae to use. One concern is the potential survival of animal pathogens within insect larvae if their feed is contaminated. In this study, the survival of a very stable virus, porcine parvovirus (PPV), within mealworm (Tenebrio molitor) and black soldier fly (BSF) (Hermetia illucens) larvae has been analyzed after oral ingestion of the virus. PPV genomic DNA could be readily detected by PCR in both species of larvae up until 9 days post ingestion (DPI), the end of the study period. Furthermore, infection of susceptible PK15 cells by PPV from homogenized mealworm larvae could be detected until at least 3 DPI, using an immunoperoxidase staining method and, up until 9 DPI, with a more sensitive real time PCR assay. Thus, PPV can remain infectious within mealworm larvae during their main growth phase through to their harvesting. However, it may be considered that PPV is exceptional in this respect since it displays unusual stability, e.g., to heat.

Cocktail vaccine for the management of Hyalomma anatolicum and Rhipicephalus microplus.

Globally, ticks rank second only to mosquitoes as vectors of deadly pathogens affecting humans and first in transmitting animal pathogens, presenting a significant challenge to human wellness and sustainability of livestock-based industries. Traditional tick control via chemical acaricides impacts on the environment and has led to the emergence of multi-acaricide-resistant tick populations. Use of immunoprophylactic, along with other components of integrated tick management, holds the potential to mitigate tick infestations in a sustainable manner. To control multi-species tick infestations, the concept of a cocktail vaccine comprising of more than one antigens has emerged as a viable solution due to the inconsistent efficacy of single antigen-based immunization protocol. In this study, a dual antigen cocktail immunization protocol was developed targeting ferritin2 (FER2) and tropomyosin (TPM) proteins, which are associated with ticks' essential cellular and physiological functions, like blood iron homeostasis and muscle contractions. Dual gene silencing of FER2 and TPM genes in Hyalomma anatolicum resulted in a 75.3% reduction in infested ticks, a 95.4% decrease in egg masses, and a complete loss of egg hatching when compared to control ticks. Microscopically, an altered ovarian cellular architecture, marked by vacuolation and reduced nucleus-to-cytoplasmic ratio were noted in the gene knocked down ticks. An immunization with cocktails of 300 µg dose of each protein, rHaFER2 and rHaTPM was standardized in a rat model and was used to immunize cross-bred (Bos indicus x B. taurus) male cattle with Montanide ISA 50V2 adjuvant on days 0, 28, and 49. A significant (p < 0.001) IgG and IgG2 antibody response was observed in the immunized animals with high IgG levels sustained until day 119 post-primary immunization, showing a 4.1-fold increase over the pre-immunization period. The animals were challenged with larvae and adults of H. anatolicum and larvae of Rhipicephalus microplus. Immunization with the cocktail antigen resulted an efficacy of 70% and 76% against H. anatolicum larvae and adults, respectively, and 54% against R. microplus infestations. Compared to single-antigen immunization, the immunization with cocktail antigens demonstrated higher protection against R. microplus and H. anatolicum ticks. The results advance the development of cocktail vaccines to control multiple tick species.

Protective Role of Indole-3-Acetic Acid Against Salmonella Typhimurium: Inflammation Moderation and Intestinal Microbiota Restoration.

Indole-3-acetic acid (IAA), a metabolite derived from microbial tryptophan metabolism, plays a crucial role in regulating intestinal homeostasis. However, the influence and potential applications of IAA in the context of animal pathogen infections remain underexplored. This study investigates the prophylactic effects of IAA pretreatment against Salmonella typhimurium (ST) SL1344 infection, focusing on its ability to attenuate inflammatory responses, enhance intestinal barrier integrity, inhibit bacterial colonization, and restore colonic microbiota dysbiosis. The results demonstrated that IAA ameliorated the clinical symptoms in mice, as evidenced by reduced weight loss and histopathological damage. Furthermore, IAA inhibited the inflammatory response by downregulating the gene expression of pro-inflammatory cytokines IL-17A, TNF-α, IL-1β, and IL-6 in colon, ileum, and liver. IAA also preserved the integrity of the intestinal mucosal barrier and promoted the expression of tight junction proteins. Additionally, 16S rRNA gene sequencing revealed significant alterations in intestinal microbiota structure induced by ST infection following IAA treatment. Notable changes in β diversity and species richness were characterized by the enrichment of beneficial bacteria including Bacteroideaceae, Spirillaceae, and Bacillus. The proliferation of Salmonella enterica subspecies enterica serovar Typhi was significantly inhibited, thereby enhancing the intestinal health of the host. In summary, the oral administration of IAA contributes to the alleviation of inflammation, restoration of the intestinal barrier, and correction of colonic microbiota disturbance in mice challenged with ST.

Vibrio alginolyticus Reprograms CIK Cell Metabolism via T3SS Effector VopS to Promote Host Cell Ferroptosis.

Vibrio alginolyticus is a Gram-negative pathogen of both marine animals and humans, resulting in significant losses for the aquaculture industry. Emerging evidence indicates that V. alginolyticus manipulates cell death for its pathogenicity, but the underlying molecular mechanisms remain unclear. Here, a gene designated vopS in V. alginolyticus HY9901 was identified, which was predicted to encode the T3SS effector protein. To determine whether VopS contributes to the pathogenesis of V. alginolyticus, the ΔvopS mutant strain was constructed and phenotypically characterized. The deletion of VopS not only reduced the ability to secrete extracellular proteases and virulence but also affected the expression of the T3SS genes. Furthermore, VopS was cytotoxic and induced apoptosis, as confirmed by elevated LDH and the activation of caspase-3. Metabolomic analysis revealed considerable metabolomic disruptions upon V. alginolyticus infection. The VopS effector induced host cell ferroptosis by promoting the synthesis of adrenic acid, depleting cellular glutathione, and subsequently increasing the accumulation of ferrous (Fe2+). Taken together, our findings provide that the VopS effector is an essential virulence factor of V. alginolyticus, which can lead to ferroptosis.

'Re-Wilding' an Animal Model With Microbiota Shifts Immunity and Stress Gene Expression During Infection.

The frequency of emerging disease is growing with ongoing human activity facilitating new host-pathogen interactions. Novel infection outcomes can also be shaped by the host microbiota. Caenorhabditis elegans nematodes experimentally colonised by a wild microbiota community and infected by the widespread animal pathogen, Staphylococcus aureus, have been shown to suffer higher mortality than those infected by the pathogen alone. Understanding the host responses to such microbiota-pathogen ecological interactions is key to pinpointing the mechanism underlying severe infection outcomes. We conducted transcriptomic analyses of C. elegans colonised by its native microbiota, S. aureus and both in combination. Correlations between altered collagen gene expression and heightened mortality in co-colonised hosts suggest the microbiota modified host resistance to infection. Furthermore, microbiota colonised hosts showed increased expression of immunity genes and variable expression of stress response genes during infection. Changes in host immunity and stress response could encompass both causes and effects of severe infection outcomes. 'Re-wilding' this model nematode host with its native microbiota indicated that typically commensal microbes can mediate molecular changes in the host that are costly when challenged by a novel emerging pathogen.