Ungulomycosis of cattle: treatment and preventive measures

Of the total number of animal diseases, 70–80 % are non-communicable diseases, and more than 50 % are surgical diseases, which cause significant economic damage to the livestock industry due to reduced productivity and premature culling of livestock. Irrational, unbalanced in terms of macro- and microelements, vitamins, carbohydrates feeding leads to metabolic disorders. At the same time, deep destructive-degenerative changes in bones, joints, ligaments and tendons are registered, horn formation in the hooves is disturbed and a less stable horn is produced. Omission in feeding unsatisfactory housing conditions and inadequate care of animals. When such conditions are often supplemented, the outer cover of animals, in particular the hooves, is affected first of all. In the event of inflammatory and dystrophic processes at the base of the skin of the hooves there is a violation of tissue (tubular) and cellular structure. The horn of such hooves undergoes changes in the form of parakeratosis and deformation; this contributes to the spread and accumulation of fungal flora, which shows its pathogenic action mainly due to enzymatic activity and toxic products, which characterizes these microorganisms as a factor in the destructive processes of the hoof horn. With reduced mechanical stability and destroyed by microscopic fungi of hoof horn is more likely to develop aseptic or purulent pododermatitis. Here, the fungal flora is not only a contributing factor in the occurrence of diseases, but also as one of the factors complicating the course of the disease. That is why timely detection and effective treatment of ungulomycosis prevents the complication of the pathological process, which can even lead to culling of animals. The essence of treatment of superficial ungulomycosis is reduced to functional cleaning of hooves and their antiseptic treatment in the form of fungicidal powders, sprays, hoof baths. In deep ungulomycosis, and, moreover, in combination with purulent pododermatitis, treatment includes local anesthesia, orthopedic treatment, antiseptic treatment with isolation, pathogenetic therapy. Prevention of ungulomycosis is based on knowledge of the etiology of the disease and the pathogenetic mechanisms of its development. Its essence is to form in animals a strong hoof horn, prevent its destruction and eliminate favorable conditions for the reproduction of keratomycetes. This includes proper feeding, proper housing conditions and appropriate animal care.

Candida gut colonization, yeast species distribution, and biofilm production in Clostridioides difficile infected patients: a comparison between three populations in two different time periods

Candida gut colonization and yeast biofilm production capacity were investigated, by means of XTT reduction assay, in Clostridioides difficile infected (CDI) patients, in non-CDI diarrheic patients, and in healthy donors in two different time periods (2013–2015 and 2018–2019 respectively). Candida gut colonization was significantly (p < 0.001) associated to C. difficile infection, and to patients infected with hypervirulent C. difficile strains bearing the tcdC deletion at nucleotide 117 (p = 0.0003). Although there was not a prevalent yeast species in CDI patients, C. albicans was the species significantly (p < 0.001) associated to both the infections sustained by the non-hypervirulent C. difficile strains and those caused by the hypervirulent strain (p = 0.001). The biofilm production by the yeasts isolated from the CDI patients and from non-CDI diarrheic patients did not differ significantly. However, a significantly (p = 0.007) higher biofilm production was observed in the Candida strains, particularly C. albicans, isolated from healthy donors compared to that of the yeasts cultured from CDI patients. Seasonal occurrence was observed in the isolation rate of CDI and non-CDI diarrheic cases (p = 0.0019), peaking in winter for CDI patients and in spring for non-CDI diarrheic patients. Furthermore, seasonality emerged in the gut colonization by Candida of CDI patients in the winter. It seems, therefore, that the reduced capacity of biofilm production by Candida strains isolated from CDI patients might have a role in the development of C. difficile infection, probably facilitating the spread of the bacteria into the gut thus amplifying their pathogenic action.

Histopathological Features of Symptomatic and Asymptomatic Honeybees Naturally Infected by Deformed Wing Virus

Deformed wing virus (DWV) is capable of infecting honeybees at every stage of development causing symptomatic and asymptomatic infections. To date, very little is known about the histopathological lesions caused by the virus. Therefore, 40 honeybee samples were randomly collected from a naturally DWV infected hive and subjected to anatomopathological examination to discriminate between symptomatic (29) and asymptomatic (11) honeybees. Subsequently, 15 honeybee samples were frozen at −80° and analyzed by PCR and RTqPCR to determinate the presence/absence of the virus and the relative viral load, while 25 honeybee samples were analyzed by histopathological techniques. Biomolecular results showed a fragment of the expected size (69bp) of DWV in all samples and the viral load was higher in symptomatic honeybees compared to the asymptomatic group. Histopathological results showed degenerative alterations of the hypopharyngeal glands (19/25) and flight muscles (6/25) in symptomatic samples while 4/25 asymptomatic samples showed an inflammatory response in the midgut and the hemocele. Results suggest a possible pathogenic action of DWV in both symptomatic and asymptomatic honeybees, and a role of the immune response in keeping under control the virus in asymptomatic individuals.

Ungulomycosis of cattle: etiological factors and pathogenetic mechanisms

Mycotic diseases, ie those caused by microscopic fungi, are widespread among animals. As for cattle, at present ungulomycosis has become the most widespread, in which keratolytic fungi affect the hooves of animals, causing complications such as deformities, pododermatitis, lesions of the hoof bone. Ungulomycosis (from the Latin ungula – hoof, Gr. Mykes – fungus) – a disease of the hooves, which are caused by microscopic fungi that have keratolytic properties. In most cases of ungulomycosis, its complications are most often registered, and not the disease itself, which does not allow to carry out effective preventive measures and treatment of animals in a timely manner. That is why the purpose of the work was to study and concretization the main factor of the disease and the contributing conditions for the disease. For this purpose the basic and additional clinical methods of research were used, including microscopy of the destroyed horn and sowing on nutrient media to determine the species content of microscopic fungi. The research was carried out in the farms of the western regions of Ukraine on cows of Black-Spotted and Holstein breeds in the stall period with tethered animals. It is found that the pathogenic action of microscopic fungi-keratomycetes on the hooves of cattle is carried out in the presence of favorable conditions of external and internal nature, primarily insufficient housing conditions and poor feeding. In particular, for osteodystrophy due to ungulomycosis, the destruction of the hoof's horn is accelerated, which complicates the pathological process. For purulent subdermatitis of traumatic origin, microscopic fungi-keratomycetes act as an additional or contributing factor to the disease. Further research will focus on the development of effective treatments and measures to prevent ungulomycosis.

Role of Electrostatic Hotspots in the Selectivity of Complement Control Proteins Toward Human and Bovine Complement Inhibition

Poxviruses are dangerous pathogens, which can cause fatal infection in unvaccinated individuals. The causative agent of smallpox in humans, variola virus, is closely related to the bovine vaccinia virus, yet the molecular basis of their selectivity is currently incompletely understood. Here, we examine the role of the electrostatics in the selectivity of the smallpox protein SPICE and vaccinia protein VCP toward the human and bovine complement protein C3b, a key component of the complement immune response. Electrostatic calculations, in-silico alanine-scan and electrostatic hotspot analysis, as introduced by Kieslich and Morikis (PLoS Comput. Biol. 2012), are used to assess the electrostatic complementarity and to identify sites resistant to local perturbation where the electrostatic potential is likely to be evolutionary conserved. The calculations suggest that the bovine C3b is electrostatically prone to selectively bind its VCP ligand. On the other hand, the human isoform of C3b exhibits a lower electrostatic complementarity toward its SPICE ligand. Yet, the human C3b displays a highly preserved electrostatic core, which suggests that this isoform could be less selective in binding different ligands like SPICE and the human Factor H. This is supported by experimental cofactor activity assays revealing that the human C3b is prone to bind both SPICE and Factor H, which exhibit diverse electrostatic properties. Additional investigations considering mutants of SPICE and VCP that revert their selectivity reveal an “electrostatic switch” into the central modules of the ligands, supporting the critical role of the electrostatics in the selectivity. Taken together, these evidences provide insights into the selectivity mechanism of the complement regulator proteins encoded by the variola and vaccinia viruses to circumvent the complement immunity and exert their pathogenic action. These fundamental aspects are valuable for the development of novel vaccines and therapeutic strategies.

Culex quinquefasciatus carrying Wolbachia is less susceptible to entomopathogenic bacteria

In an attempt to evaluate the susceptibility of the mosquito Culex quinquefasciatus to bacterial agents, a population naturally infected with a Wolbachia pipientis wPipSJ native strain was tested against the action of three bacterial mosquitocides, Bacillus thuringiensis subsp. israelensis, Bacillus wiedmannii biovar thuringiensis and Lysinibacillus sphaericus. Tests were carried out on mosquito larvae with and without Wolbachia (controls). Cx. quinquefasciatus naturally infected with the native wPipSJ strain proved to be more resistant to the pathogenic action of the three mosquitocidal bacterial strains. Additionally, wPipSJ was fully characterised using metagenome-assembled genomics, PCR–RFLP (PCR-Restriction Fragment Length Polymorphism) and MLST (MultiLocus Sequence Typing) analyses. This Wolbachia strain wPipSJ belongs to haplotype I, group wPip-III and supergroup B, clustering with other mosquito wPip strains, such as wPip PEL, wPip JHB, wPip Mol, and wAlbB; showing the southernmost distribution in America. The cytoplasmic incompatibility phenotype of this strain was revealed via crosses between wildtype (Wolbachia+) and antibiotic treated mosquito populations. The results of the tests with the bacterial agents suggest that Cx. quinquefasciatus naturally infected with wPipSJ is less susceptible to the pathogenic action of mosquitocidal bacterial strains when compared with the antibiotic-treated mosquito isoline, and is more susceptible to B. thuringiensis subsp. israelensis than to the other two mosquitocidal agents.

Molecular Targeting of VEGF with a Suramin Fragment–DOCA Conjugate by Mimicking the Action of Low Molecular Weight Heparins

Molecular targeting of growth factors has shown great therapeutic potential in pharmaceutical research due to their roles in pathological conditions. In the present study, we developed a novel suramin fragment and deoxycholic acid conjugate (SFD) that exhibited the potential to bind to the heparin-binding site (HBD) of vascular endothelial growth factor (VEGF) and to inhibit its pathogenic action for the first time. Notably, SFD was optimally designed for binding to the HBD of VEGF using the naphthalenetrisulfonate group, allowing to observe its excellent binding efficacy in a surface plasmon resonance (SPR) study, showing remarkable binding affinity (KD = 3.8 nM) as a small molecule inhibitor. In the tubular formation assay, it was observed that SFD could bind to HBD and exhibit antiangiogenic efficacy by inhibiting VEGF, such as heparins. The cellular treatment of SFD resulted in VEGF-inhibitory effects in human umbilical vein endothelial cells (HUVECs). Therefore, we propose that SFD can be employed as a novel drug candidate to inhibit the pathophysiological action of VEGF in diseases. Consequently, SFD, which has a molecular structure optimized for binding to HBD, is put forward as a new chemical VEGF inhibitor.

Design and Synthesis of Small Molecules as Potent Staphylococcus aureus Sortase A Inhibitors

The widespread and uncontrollable emergence of antibiotic-resistant bacteria, especially methicillin-resistant Staphylococcus aureus, has promoted a wave of efforts to discover a new generation of antibiotics that prevent or treat bacterial infections neither as bactericides nor bacteriostats. Due to its crucial role in virulence and its nonessentiality in bacterial survival, sortase A has been considered as a great target for new antibiotics. Sortase A inhibitors have emerged as promising alternative antivirulence agents against bacteria. Herein, the structural and preparative aspects of some small synthetic organic compounds that block the pathogenic action of sortase A have been described.

Genomic and Proteomic Comparative Analysis of SARS-CoV-2 versus SARS-CoV-GD01

Background Since the emergence of the pandemic novel pneumonia (COVID-19) disease in Wuhan city in China in November 2019, it is becoming holistically urgent to discover and definitely determine the potential origin of causative virus of this disease, SARS-CoV2 to understand its pathogenic action an better design proper remedies. Methods Using bioinformatics analysis, the whole genome of SARS-CoV2 emerging in 2020 and its deduced proteome were compared with the corresponding information on SARS-CoV-GD01 having emerged in 2003 in China. The genomes squences of the two viruses were obtained from NCBI. Alignment of protein sequences for all genes of both genomes were performed and displayed using Clustal Omega data base. Results Bioinformatics analysis revealed 10 genes encoding 10 proteins in the SARS-CoV2 genome instead of 11 genes encoding 12 proteins in the case of SARS-CoV-GD01, where the first gene is uniquely encoding two glycoproteins. Additionally, bio-informatics analysis disclosed variations in SARS-CoV2 genome size as a result of nucleotides insertion and deletion in all genes of the virus especially orf1ab gene, spike gene, and ORF10 gene. The most conspicuous alteration is apparently noticed in the spike gene, encoding for a novel protein enabling the virus to attach to the cell membrane via the interaction with host cell receptor, initiating probably a new pathway of infection and a specific pathogenic action. This alteration is Conclusions The big alterations in the genome of SARS-CoV-2 from that of SARS-CoV-GD01 may be potentially responsible for the worldwide witnessed high virulence and accelerated spread. The qualified and quantified information presented in the current study on the SARS-CoV-2, detailing the specificity and the magnitude of genomic and proteomic alterations from SARS-CoV-GD01, developed probably during 16 years will not only enable designing right drugs and strategies of confronting the current viral version, but it may rather allow to extrapolate and foresee potential outbreaks of newer versions during the coming decades. At the time of epidemics, nonspecific ways and drugs should be resorted to for confronting emergent viral infections. Chemically modified positively charged proteins and peptides can offer a wealth of potential antiviral agents but need more clinical research.