Biodegradable Polymers in Veterinary Medicine-A Review.

During the past two decades, tremendous progress has been made in the development of biodegradable polymeric materials for various industrial applications, including human and veterinary medicine. They are promising alternatives to commonly used non-degradable polymers to combat the global plastic waste crisis. Among biodegradable polymers used, or potentially applicable to, veterinary medicine are natural polysaccharides, such as chitin, chitosan, and cellulose as well as various polyesters, including poly(ε-caprolactone), polylactic acid, poly(lactic-co-glycolic acid), and polyhydroxyalkanoates produced by bacteria. They can be used as implants, drug carriers, or biomaterials in tissue engineering and wound management. Their use in veterinary practice depends on their biocompatibility, inertness to living tissue, mechanical resistance, and sorption characteristics. They must be designed specifically to fit their purpose, whether it be: (1) facilitating new tissue growth and allowing for controlled interactions with living cells or cell-growth factors, (2) having mechanical properties that address functionality when applied as implants, or (3) having controlled degradability to deliver drugs to their targeted location when applied as drug-delivery vehicles. This paper aims to present recent developments in the research on biodegradable polymers in veterinary medicine and highlight the challenges and future perspectives in this area.

Cleft Lip and Palate in Four Full-Sib Puppies from a Single Litter of Staffordshire Bull Terrier Dogs: An Anatomical and Genetic Study.

Cleft lip and palate (CLP) is a well-known congenital defect in dogs, characterized by abnormal communication between the oral and nasal cavities. Its incidence rate is high and affects all dog breeds. The etiology of CLP is thought to be multifactorial, caused by both genetic and environmental factors. In this study, four puppies out of seven from a single litter of Staffordshire Bull Terrier dogs with craniofacial abnormalities were anatomically and genetically examined. Classical anatomical preparation, dyed-latex-injection of the arterial vessels, and cone-beam computed tomography were used. The puppies showed variations in their observable abnormalities: three of them had a complete cleft of the palate on both sides, while one puppy had a cleft on the right side only. Cytogenetic analysis showed a normal diploid chromosome number (2n = 78,XX or 78,XY) in the studied animals. Known genomic variants of CLP were examined in the ADAMTS20, DLX6, and MYH3 genes, but no mutations were identified. Further studies are needed to identify the breed-specific genetic variants associated with canine CLP.

The Shape of the Nasal Cavity and Adaptations to Sniffing in the Dog (Canis familiaris) Compared to Other Domesticated Mammals: A Review Article.

Dogs are a good starting point for the description and anatomical analysis of turbinates of the nose. This work aimed at summing up the state of knowledge on the shape of the nasal cavity and airflow in these domestic animals and dealt with the brachycephalic syndrome (BOAS) and anatomical changes in the initial airway area in dogs with a short and widened skull. As a result of artificial selection and breeding concepts, the dog population grew very quickly. Modern dog breeds are characterized by a great variety of their anatomical shape. Craniological changes also had a significant impact on the structure and physiology of the respiratory system in mammals. The shape of the nasal cavity is particularly distinctive in dogs. Numerous studies have established that dogs and their olfactory ability are of great importance in searching for lost people, detecting explosives or drugs as well as signaling disease in the human body. The manuscript describes the structure of the initial part of the respiratory system, including the nasal turbinates, and compares representatives of various animal species. It provides information on the anatomy of brachycephalic dogs and BOAS. The studies suggest that further characterization and studies of nasal turbinates and their hypertrophy are important.

Congenital Malformations in a Holstein-Fresian Calf with a Unique Mosaic Karyotype: A Case Report.

A Holstein-Fresian calf with multiple congenital malformations was subjected postmortem to anatomical and genetic investigation. The calf was small (20 kg), had shortened limbs and was unable to stand up. It lived only 44 days. Detailed anatomical investigation revealed the following features: head asymmetry, the relocation of the frontal sinus and eye orbits, hypoplastic thymus without neck part, ductus Botalli, unfinished obliteration in umbilical arteries, and a bilateral series of tooth germs in the temporal region. Cytogenetic examination, performed on in vitro cultured fibroblasts, showed a unique mosaic karyotype with a marker chromosome-60,XX[9 2%]/60,XX,+mar[8%], which was for the first time described in cattle. No other chromosome abnormalities indicating chromosome instabilities, like chromatid breaks or gaps were identified, thus teratogenic agent exposure during pregnancy was excluded. The marker chromosome (mar) was small and it was not possible to identify its origin, however, sequential DAPI/C (4',6-diamidino-2-phenylindole) band staining revealed a large block of constitutive heterochromatin, which is characteristic for centromeric regions of bovine autosomes. We suppose that the identified marker chromosome was a result of somatic deletion in an autosome and its presence could be responsible for the observed developmental malformations. In spite of the topographic distance among the affected organs, we expected a relationship between anatomical abnormalities. To the of our best knowledge, this is the first case of a mosaic karyotype with a cell line carrying a small marker chromosome described in a malformed calf.

Comparative Evaluation of Piper nigrum, Rosmarinus officinalis, Cymbopogon citratus and Juniperus communis L. Essential Oils of Different Origin as Functional Antimicrobials in Foods.

Essential oils can be used as preservatives in foods because of their ability to inhibit bacteria growth in low concentration, which does not influence on foods' organoleptic properties and does not generate the resistance mechanisms in cells. The aim of that work was to compare the effectiveness of commercial oils from black pepper (Piper nigrum), rosemary (Rosmarinus officinalis), lemongrass (Cymbopogon citratus) and juniper (Juniperus communis L.) with oils obtained in our laboratory. The typical cultivation method was supported by the flow cytometry to detect the cells of very low physiologic and metabolic activity. Our investigation demonstrated that both types of oils can effectively inhibit the growth of saprophytic bacteria P. orientalis. The oils distilled in our laboratory had a bacteriostatic effect at a lower concentration, which is important for application in the food industry. Flow cytometry analyzes and confirmed the thesis that essential oils do not have a germicidal effect on bacteria cells.

The Influence of Selected Plant Essential Oils on Morphological and Physiological Characteristics in Pseudomonas Orientalis.

The aim of this work was to estimate the antibacterial activity of selected essential oils on Pseudomonas orientalis strains isolated from foods. An attempt was also made to identify the mechanisms of the action of the plant oils. Classical methods of assessment of the effectiveness of antimicrobial activity of oils were linked with flow cytometry. It was observed that bergamot, lemongrass, bitter orange, juniper, and black pepper oils have bacteriostatic effect against P. orientalis P49. P. orientalis P110 is sensitive to lime, lemongrass, juniper, rosemary, and black pepper oils. Additionally, plant oils with biostatic effect on P. orientalis limited the intracellular metabolic activity of cells; this was closely linked with the ability of plant oils' bioactive components to interact with bacteria cell membrane, causing the release of membrane proteins. As a result, the selective permeability of the cell membranes were damaged and the bacterial shape was transformed to coccoid in form.

The industrial potential of herbs and spices - a mini review.

Herbs and spices have been used for food and medicinal purposes for centuries - the first recorded evidence of their use dates back to 1500BC and the Ebers Papyrus, which mentioned spices such as anise, mustard, saffron, cinnamon, and cassia. Now, in the 21st century, a variety of secondary compounds produced by plants are used in many fields of industry, such as food production (to improve taste, to provide vitamins and macro- and microelements, and also to inhibit food spoilage caused by foodborne bacteria), in medicine (in the treatment of various diseases; in chemoprevention and cancer therapy; as a source of natural antimicrobials for the treatment of infectious disease), and in pharmacology and cosmetology (in dietary supplements, and as a result of the demand for preservative-free cosmetics, to reduce the risk of methylparaben allergies). The aim of this review is to present the major active compounds in herbs and spices and explore their potential applications in industry.

Role of gallic and p-coumaric acids in the AHL-dependent expression of flgA gene and in the process of biofilm formation in food-associated Pseudomonas fluorescens KM120.

BACKGROUND: In the process of Pseudomonas fluorescens biofilm formation, N-acyl-l-homoserine lactone (AHL)-mediated flagella synthesis plays a key role. Inhibition of AHL production may attenuate P. fluorescens biofilm on solid surfaces. This work validated the anti-biofilm properties of p-coumaric and gallic acids via the ability of phenolics to suppress AHL synthesis in P. fluorescens KM120. The dependence between synthesis of AHL molecules, expression of flagella gene (flgA) and the ability of biofilm formation by P. fluorescens KM120 on a stainless steel surface (type 304L) was also investigated. RESULTS: Research was carried out in a purpose-built flow cell device. Limitations on AHL synthesis in P. fluorescens KM120 were observed at concentrations of 120 and 240 µmol L(-1) of phenolic acids in medium. At such levels of gallic and p-coumaric acids the ability of P. fluorescens KM120 to synthesize 3-oxo-C6-homoserine lactone (HSL) was not observed. These concentrations caused decreased expression of flgA gene in P. fluorescens KM120. The changes in expression of AHL-dependent flgA gene significantly decreased the rate of microorganism colonization on the stainless steel surface. CONCLUSION: Phenolic acids are able to inhibit biofilm formation. The results obtained in the work may help to develop alternative techniques for anti-biofilm treatment in the food industry. © 2015 Society of Chemical Industry.

Biotechnological potential of Clostridium butyricum bacteria.

In response to demand from industry for microorganisms with auspicious biotechnological potential, a worldwide interest has developed in bacteria and fungi isolation. Microorganisms of interesting metabolic properties include non-pathogenic bacteria of the genus Clostridium, particularly C. acetobutylicum, C. butyricum and C. pasteurianum. A well-known property of C. butyricum is their ability to produce butyric acid, as well as effectively convert glycerol to 1,3-propanediol (38.2 g/L). A conversion rate of 0.66 mol 1,3-propanediol/mol of glycerol has been obtained. Results of the studies described in the present paper broaden our knowledge of characteristic features of C. butyricum specific isolates in terms of their phylogenetic affiliation, fermentation capacity and antibacterial properties.

Hypothetical glycerol pathways of newly isolated strains capable of 1,3-propanediol production.

Study presented here demonstrates the ability of three newly isolated strains, obtained from environmental probes (manure, bottom sediment, and food waste) and identified as Clostridium bifermentans, Clostridium butyricum, and Hafnia alvei, to synthesize 1,3-propanediol (1,3-PD), organic acids (such as lactic, acetic, fumaric, succinic, and butyric acids), and ethanol from glycerol. The production of 1,3-PD as well as the glycerol pathways in C. bifermentans and H. alvei cells have not been investigated and described yet by others. Moreover, there is no data in the available literature on the products of glycerol utilization by H. alvei and there is only some incoherent data (mainly from the first half of the twentieth century) about the ability of C. bifermentans to carry out glycerol degradation. Additionally, this study presents complete hypothetical glycerol pathways and the basic fermentation kinetic parameters (such as yield and productivity) for both strains as well as for the newly isolated C. butyricum strain.

Biotechnological potential of Clostridium butyricum bacteria

In response to demand from industry for microorganisms with auspicious biotechnological potential, a worldwide interest has developed in bacteria and fungi isolation. Microorganisms of interesting metabolic properties include non-pathogenic bacteria of the genus Clostridium, particularly C. acetobutylicum, C. butyricum and C. pasteurianum. A well-known property of C. butyricum is their ability to produce butyric acid, as well as effectively convert glycerol to 1,3-propanediol (38.2 g/L). A conversion rate of 0.66 mol 1,3-propanediol/mol of glycerol has been obtained. Results of the studies described in the present paper broaden our knowledge of characteristic features of C. butyricum specific isolates in terms of their phylogenetic affiliation, fermentation capacity and antibacterial properties.

Impurities of crude glycerol and their effect on metabolite production.

Glycerol is a valuable raw material for the production of industrially useful metabolites. Among many promising applications for the use of glycerol is its bioconversion to high value-added compounds, such as 1,3-propanediol (1,3-PD), succinate, ethanol, propionate, and hydrogen, through microbial fermentation. Another method of waste material utilization is the application of crude glycerol in blends with other wastes (e.g., tomato waste hydrolysate). However, crude glycerol, a by-product of biodiesel production, has many impurities which can limit the yield of metabolites. In this mini-review we summarize the effects of crude glycerol impurities on various microbial fermentations and give an overview of the metabolites that can be synthesized by a number of prokaryotic and eukaryotic microorganisms when cultivated on glycerol.

Influence of phenolic acids on indole acetic acid production and on the type III secretion system gene transcription in food-associated Pseudomonas fluorescens KM05.

The purpose of these investigations was to evaluate the reduction capability of phenolic acids (ferulic, chlorogenic, gallic, and p-coumaric acids) on indole acetic acid synthesis by food-associated Pseudomonas fluorescens KM05. Specific genetic primer for the type III secretion system (TTSS) in P. fluorescens KM05 was designed and the influence of phenolic acids on its expression was investigated. In the work the ferulic and chlorogenic acids at the concentration of 0.02 and 0.04 µg/ml affected on bacterial growth pattern and the signal molecules production. The phenolic acids, that were appreciable effective against P. fluorescens KM05 indole acetic acid production, significantly suppressed TTSS gene.

An increasing of the efficiency of microbiological synthesis of 1, 3-propanediol from crude glycerol by the concentration of biomass

Background 1,3-Propanodiol (1,3-PD), is used in the production of polytrimethylene terephthalate (PTT), an aromatic polyester that exhibits high elastic recoveries. It is also employed as a supplement with low solidification properties, a solvent and a lubricant in the formof propylene glycol. 1,3-PD is effectively synthesized by a microbiological way from crude glycerol. The main problem of this technology is using a high concentration of glycerol, which is a limiting factor for bacteria cells growth (especially in batch fermentation). Results In this work, the influence of different glycerol concentration in batch fermentation on Clostridium butyricum DSP1 metabolism was investigated. The biomass was concentrated for two times with the use of membrane module (in case of increasing kinetic parameters). Increased optical density of bacteria cells six times increased the productivity of 1,3-PD in cultivation with 20 g/L of glycerol at the beginning of the process, and more than two times in cultivation with 60-80 g/L. Also the possibility of complete attenuation of 140 g/L of crude glycerol in the batch fermentation was investigated. During the cultivation, changes of protein profiles were analyzed. The most significant changes were observed in the cultivation in the medium supplemented with 80 g/L of glycerol. They related mainly to the DNA protein reconstructive systems, protective proteins (HSP), and also the enzymatic catalysts connected with glycerol metabolic pathway. Conclusions The application of filtration module in batch fermentation of crude glycerol by C. butyricum DSP1 significantly increased the productivity of the process.

Microbial purification of postfermentation medium after 1,3-PD production from raw glycerol.

1,3-Propanediol (1,3-PD) is an important chemical product which can be used to produce polyesters, polyether, and polyurethanes. In the process of conversion of glycerol to 1,3-PD by Clostridium large number of byproducts (butyric, acetic and lactic acid) are generated in the fermentation medium. The aim of this work was to isolate bacteria strains capable of the utilization of these byproducts. Screening of 30 bacterial strains was performed using organic acids as carbon source. Selected isolates were taxonomically characterized and identified as Alcaligenes faecalis and Bacillus licheniformis. The most active strains, Alcaligenes faecalis JP1 and Bacillus licheniformis JP19, were able to utilize organic acids almost totally. Finally, it was find out that by the use of coculture (C. butyricum DSP1 and A. faecalis JP1) increased volumetric productivity of 1,3-PD production (1.07 g/L/h) and the yield equal to 0.53 g/g were obtained in bioreactor fermentation. Moreover, the only by-product present was butyric acid in a concentration below 1 g/L.

The ability of Clostridium bifermentans strains to lactic acid biosynthesis in various environmental conditions.

Clostridium bifermentans strains, isolated from a manure, were examinated for their ability to produce lactic acid from PY medium with glycerol under different pH conditions and when PY medium was supplemented with saccharides such as fructose, sorbitol, glucose, mannose, mannitol, maltose, xylose, raffinose, and arabinose. In the last test performed, the ability of investigated strains to produce lactic acid from mixed carbon source (glycerol plus saccharide) was checked. The strains of Cl. bifermentans, designated as CB 371, CB 374, and CB 376 grew and produced lactic acid on PY medium irrespective of pH and the carbon source used. The optimal lactic acid production on PY medium with glycerol was obtained at pH of 7.0 in case of CB 371 and 376 (19.63 g/L and 16.65 g/L, accordingly) and at pH 8.0 in case of CB 374 (13.88 g/L). The best productivity of lactic acid on PY media by CB 371, CB 374, and CB 376 (above 30 g/L) was observed when mannitol was used as a carbon source. The mixed carbon source did not increase productivity of lactic acid by Cl. bifermentans. The yield of lactic acid was approximately equal to the yield of lactic acid obtained on the medium with only glycerol and lower than in medium with only mannitol. Thus, from the environmental point of view it is more beneficial to use the medium with waste-type material only, such as glycerol.

Beneficial and harmful roles of bacteria from the Clostridium genus.

Bacteria of the Clostridium genus are often described only as a biological threat and a foe of mankind. However, many of them have positive properties and thanks to them they may be used in many industry branches (e.g., in solvents and alcohol production, in medicine, and also in esthetic cosmetology). During the last 10 years interest in application of C. botulinum and C. tetani in medicine significantly increased. Currently, the structure and biochemical properties of neurotoxins produced by these bacterial species, as well as possibilities of application of such toxins as botulinum as a therapeutic factor in humans, are being intensely researched. The main aim of this article is to demonstrate that bacteria from Clostridium spp. are not only pathogens and the enemy of humanity but they also have many important beneficial properties which make them usable among many chemical, medical, and cosmetic applications.

Isolation process of industrially useful Clostridium bifermentans from natural samples.

A selective isolation procedure of clostridial strains from natural samples able to convert glycerol to 1,3-propanediol (1,3-PD) and organic acids was investigated. The modified PY medium of high concentration of NaHCO(3) was shown to be highly selective for Clostridium bifermentans. Obtained isolates produced mainly 1,3-PD, lactic, acetic, and formic acids from glycerol.

Application of fast liquid chromatography for antioxidants analysis.

BACKGROUND: An intensive development of the Fast Liquid Chromatography (FLC) has been recently observed. It makes possible to reduce time analysis and improve resolution as well as sensitivity. The aim of this study was to separate the chosen antioxidants optimization using the FLC method. MATERIAL AND METHODS: The three various procedures for antioxidants analysis were compared. Mobile phases containing aqueous solution of formic acid, acetic acid, acetonitrile, and methanol were tested. Limit of detection (LOD), limit of quantification (LOQ), linearity and repeatability of each procedures were determined. RESULTS: Developed procedure enabled to separate all analytes and allowed to get low LOD levels and good repeatability. This procedure was used for antioxidants analysis in buckwheat and buckwheat products. CONCLUSION: Fast Liquid Chromatography allows to reduce time analysis and obtain good validation parameters.