Inclusion of a Bacillus-based probiotic in non-starch polysaccharides-rich broiler diets.

This study examined the effects of a 3-strain Bacillus-based probiotic (BP; Bacillus amyloliquefaciens and two Bacillus subtilis) in broiler diets with different rye levels on performance, mucus, viscosity, and nutrient digestibility. We distributed 720 one-d-old female broilers into 72 pens and designed nine diets using a 3 × 3 factorial approach, varying BP levels (0, 1.2 × 106, and 1.2 × 107 CFU/g) and rye concentrations (0, 200, 400 g/kg). On d 35, diets with 200 or 400 g/kg rye reduced broiler weight gain (BWG). Diets with 400 g/kg rye had the highest FCR, while rye-free diets had the lowest (p ≤0.05). Adding BP increased feed intake and BWG in weeks two and three (p ≤0.05). It should be noted that the overall performance fell below the goals of the breed. Including rye in diets reduced the coefficient of apparent ileal digestibility (CAID) for protein, ether extract (EE), calcium, phosphorus, and all amino acids (p ≤0.05). Rye-free diets exhibited the highest CAID for all nutrients, except for methionine, EE, and calcium, while diets with 400 g/kg of rye demonstrated the lowest CAID (p ≤0.05). BP in diets decreased phosphorus CAID (p ≤0.05). Diets containing 1.2 × 107 CFU/g (10X) of BP exhibited higher CAID of methionine than the other two diets (p ≤0.05). Diets containing 10X of BP showed higher CAID of cysteine than diets with no BP (p ≤0.05). Ileal viscosity increased as the inclusion level of rye in the diets increased (p ≤0.05). The ileal concentration of glucosamine in chickens fed diets with 400 g/kg of rye was higher than in those fed diets with no rye (p ≤0.05). Furthermore, ileal galactosamine concentrations were elevated in diets with 200 and 400 g/kg of rye when compared to rye-free diets (p ≤0.05). However, BP in diets had no impact on ileal viscosity, galactosamine, or glucosamine (p > 0.05). In conclusion, the applied Bacillus strains appeared to have a limited capacity to produce arabinoxylan-degrading enzymes and were only partially effective in mitigating the negative impacts of rye arabinoxylans on broilers.

Impact of feed additives and host-related factors on bacterial metabolites, mucosal integrity and immune response in the ileum of broilers.

The present study aimed to investigate the effect of age, breed, and sex of broilers, as well as a probiotic or phytobiotic product on mucosal morphology, bacterial metabolites, and immune traits in the ileum of broilers. A total of 2,880 one-day-old male and female broiler chicks from two breeds (Ross308® and Cobb500®) were randomly assigned to 72 pens. Broilers were offered a wheat-soybean diet without (CO), or with either a probiotic (PO; 2.4 × 109 CFU/kg of Bacillus subtilis DSM32324 and DSM32325 and B. amyloliquefaciens DSM25840) or a phytobiotic (PY; grape extract, 165 ppm procyanidin and 585 ppm polyphenols of the diet) product. The trial was conducted with a 3 × 2 × 2 factorial arrangement of diet, breed, and sex in a completely randomized design (6 replicate-pens per treatment). At day 7, 21, and 35, one chicken per pen was slaughtered for collecting ileal tissue to evaluate of histomorphology and mRNA expression, as well as ileal digesta to measure bacterial metabolites. Data were subjected to ANOVA (the main factors; age, diet, breed, and sex) and Four-Way ANOVA (interactions) using GLM procedure. Overall, the concentration of acetate and total short chain fatty acids reached the peak and lactate decreased to its lowest on day 21, but their concentrations at day 7 and 35 were similar (p > 0.05). Spermine, spermidine, and ammonia decreased after day 7, while putrescine and cadaverine increased after day 21 (p < 0.05). mRNA expression of cytokines, mucin 2 (MUC2) and claudin 5 (CLDN5) was similar; increased from day 7 to 21 and decreased afterward (p < 0.05). Villus height, crypt depth and villus surface area increased with age (p < 0.05). Acidic goblet cells (GC) number and density increased after day 21 (p < 0.05). Ross broilers showed higher D-lactate concentration and IFN-γ expression, while Cobb broilers had greater IL-4, IL-6 and TNF-α expression and higher total GC number (p < 0.05). Female displayed higher villus height and GC number and density (mixed and total GC) than male (p < 0.05). The effect of dietary treatment was not found on any investigated variables (p > 0.05). In conclusion, aging of broilers affected ileal histomorphology, cytokine expression, and barrier integrity, as well as bacterial activity. These observed impacts could be attributed to host-microbiota interaction and the direct effects of bacterial metabolites on intestinal cells and immune system.

Alterations in bacterial metabolites, cytokines, and mucosal integrity in the caecum of broilers caused by feed additives and host-related factors.

A total of 2,880 one-day-old male and female broiler chicks from two breeds, Ross308 and Cobb500 were randomly assigned to 72 pens. Broilers were offered three diets: a wheat-soybean diet without (CO), or with either a probiotic (probiotic; 2.4 x 109 CFU/kg diet of Bacillus subtilis DSM32324 and DSM32325 and B. amyloliquefaciens DSM25840) or a phytobiotic (phytobiotic; grape extract with 165 ppm procyanidin and 585 ppm polyphenol) product. The trial was conducted with a 3 × 2 × 2 factorial arrangement of diet, breed and sex in a completely randomized design and consisted of 6 replicate-pens per treatment (40 birds per pen). At day 7, 21, and 35, one chicken per pen was slaughtered for caecal sampling to quantify bacterial metabolites (digesta) as well as evaluate mRNA abundance and histomorphology (tissue). Data were subjected to ANOVA using GLM procedure to evaluate age, diet, breed and sex and their interactions. Spearman's correlation (r) was analyzed between metabolite concentration and mRNA abundance. Overall, the concentration of short chain fatty acids increased with age, while lactate decreased from day 7 to 21 (p < 0.05). The mRNA abundance of IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-17α, IL-18, IFN-γ and TGF-ß2 increased with age but IL-1ß and TNF-α increased in abundance from day 7 to 21 and then decreased (p < 0.05). Abundance of MUC2 and CLDN5 increased after day 21 (p < 0.05). Caecal crypt depth increased with age (p < 0.05). Acidic goblet cell (GC) number peaked at day 21 (p < 0.05), while mixed GC number was not affected by age. A few impacts of breed, diet and interactions on the investigated variables showed no meaningful biological pattern. Propionate positively correlated with all cytokines investigated (r = 0.150-0.548), except TNF-α. Lactate negatively correlated with pro-inflammatory cytokines like IL-1ß (r = -0.324). Aging affected caecal histomorphology, bacterial activity and genes responsible for barrier integrity and inflammatory response. This effect could be attributed to the interaction between gut microbiota and immune system as well as the direct effect of metabolites on gut histomorphology and cytokine mRNA abundance.

Effects of potential probiotics on the expression of cytokines and human ß-defensins in human gingival epithelial cells and in vivo efficacy in a dog model.

OBJECTIVES: The aims of this study were to examine the effects of probiotic strains on the expression of cytokines and human ß-defensins-2-4 in human gingival epithelial cells and to investigate the in vivo efficacy of probiotics in a dog model. DESIGN: Selected probiotics, Lacticaseibacillus paracasei SD1, L. rhamnosus SD4, L. rhamnosus SD11 and Limosilactobacillus fermentum SD7, were examined for the expression of human ß-defensins-2-4, and cytokine responses after stimulated by various periodontopathogens. Subsequently, an in vivo study was set as a randomized, double-blind, placebo-controlled intervention in a dog model. A total of 20 dogs with mild gingivitis were randomly assigned to either the probiotic or control group. The effects of probiotics on periodontopathogenic- and cytokine levels were analyzed after 4 weeks of intervention. RESULTS: It showed that all probiotics could induce human ß-defensins-2-4, interleukin-1ß, interleukin-6, interleukin-8, and tumor necrosis factor-α expressions in human gingival epithelial cells; however, a significant difference had been found among the strains. When individual probiotic strain was combined to periodontopathogens, a significant reduction of IL-8 expression was found. Results of the in vivo study demonstrated that the bacterial levels, especially Porphyromonas gingivalis, and the interleukin-8 levels were significantly decreased after receiving the probiotic products compared to the controls. CONCLUSION: The results of the in vitro and in vivo studies in dogs are encouraging to support the effectiveness of a mixture of probiotic treatments for improvement of periodontal health by reducing periodontopathogens and interleukin-8 levels. Therefore, such probiotics preparation could possibly have therapeutic potential in human periodontitis.

Development and Functional Properties of Intestinal Mucus Layer in Poultry.

Intestinal mucus plays important roles in protecting the epithelial surfaces against pathogens, supporting the colonization with commensal bacteria, maintaining an appropriate environment for digestion, as well as facilitating nutrient transport from the lumen to the underlying epithelium. The mucus layer in the poultry gut is produced and preserved by mucin-secreting goblet cells that rapidly develop and mature after hatch as a response to external stimuli including environmental factors, intestinal microbiota as well as dietary factors. The ontogenetic development of goblet cells affects the mucin composition and secretion, causing an alteration in the physicochemical properties of the mucus layer. The intestinal mucus prevents the invasion of pathogens to the epithelium by its antibacterial properties (e.g. ß-defensin, lysozyme, avidin and IgA) and creates a physical barrier with the ability to protect the epithelium from pathogens. Mucosal barrier is the first line of innate defense in the gastrointestinal tract. This barrier has a selective permeability that allows small particles and nutrients passing through. The structural components and functional properties of mucins have been reviewed extensively in humans and rodents, but it seems to be neglected in poultry. This review discusses the impact of age on development of goblet cells and their mucus production with relevance for the functional characteristics of mucus layer and its protective mechanism in the chicken's intestine. Dietary factors directly and indirectly (through modification of the gut bacteria and their metabolic activities) affect goblet cell proliferation and differentiation and can be used to manipulate mucosal integrity and dynamic. However, the mode of action and mechanisms behind these effects need to be studied further. As mucins resist to digestion processes, the sloughed mucins can be utilized by bacteria in the lower part of the gut and are considered as endogenous loss of protein and energy to animal. Hydrothermal processing of poultry feed may reduce this loss by reduction in mucus shedding into the lumen. Given the significance of this loss and the lack of precise data, this matter needs to be carefully investigated in the future and the nutritional strategies reducing this loss have to be defined better.

Coronal Alignment Correction and Maintenance of Tibial Slope in Opening-Wedge Valgus High Tibial Osteotomy Using a 4-Reference Kirschner Wire Technique: A Cadaveric Study.

BACKGROUND: Opening-wedge valgus high tibial osteotomy (OWHTO) is a common surgical procedure used to treat symptomatic varus femorotibial malalignment in adults. Several intraoperative methods are available to determine the correct correction alignment, but achieving the desired alignment correction is difficult. PURPOSE/HYPOTHESIS: The aim of this study was to assess a 4-reference K-wire technique that is relatively easy to apply and can reliably assess actual alignment correction during surgery after determination of the desired corrective angle. We hypothesized that this technique would accurately determine the coronal correction and properly maintain the tibial slope intraoperatively during OWHTO. STUDY DESIGN: Descriptive laboratory study. METHODS: This study was conducted using 12 fresh-frozen cadavers; 12 randomly chosen knees were corrected 5° and 12 knees were corrected 10° by use of 2 coronal and 2 sagittal K-wires. The first and second coronal K-wires were drilled at 4 cm and 1 to 2 cm below the medial joint line toward the tibiofibular joint, respectively. The angles of these 2 coronal K-wires were measured before and after the gap was opened via a modified goniometer. The difference in the angle formed by these 2 coronal K-wires from before to after opening of the gap was the alignment correction angle. In addition, 2 sagittal K-wires were drilled parallel to each other before the gap opening above and below the osteotomy site. Ensuring that these 2 sagittal K-wires remained parallel after the gap opening confirmed that the tibial slope had been maintained. The paired t test was used to compare the desired alignment corrections and the different angles measured between the pre- and postoperative radiographic alignments. RESULTS: The mean ± SD differences in angles between the pre- and postoperative alignments of the 5° and 10° corrections were 5.04° ± 0.68° and 10.03° ± 0.68°, respectively, indicating no statistically significant differences between pre- and postoperative alignment in both groups. As well, no significant difference was noted between the pre- and postoperative medial tibial slope (P = .54). CONCLUSION: The coronal alignment correction and maintenance of the tibial slope using the 4-reference K-wire technique was found to be highly accurate and reliable. CLINICAL RELEVANCE: Achieving the correct angle in OWHTO is difficult, and the 4-reference K-wire technique provides an easier and more reliable way to obtain the correct angle. This technique can be used in most hospital settings, with no need for expensive equipment.

Coracoclavicular Stabilization With Two Loops of Equal Tension Using a Double O Loops Technique in the Distal Clavicle Fracture.

The distal clavicle fracture is one of the most common injuries around the shoulder joint. There is no consensus regarding a gold standard treatment. Each technique has advantages and disadvantages. Currently, coracoclavicular (CC) stabilization is one of the most popular techniques because this operative procedure provides good stability of the fracture and has few complications. The CC stabilization is a suspensory fixation that consists of many two-CC-loop arrangements. It is, however, difficult to gain equal tension in both CC loops because one loop is always tighter and has greater action in maintaining bone alignment than the other loop. To solve this problem, we propose a double O loops technique to achieve two equal tension loops.

Patient Position Is Related to the Risk of Neurovascular Injury in Clavicular Plating: A Cadaveric Study.

BACKGROUND: Fixation of clavicle shaft fractures with a plate and screws can endanger the neurovascular structures if proper care is not taken. Although prior studies have looked at the risk of clavicular plates and screws (for example, length and positions) to vulnerable neurovascular structures (such as the subclavian vein, subclavian artery, and brachial plexus) in the supine position, no studies to our knowledge have compared these distances in the beach chair position. QUESTIONS/PURPOSES: (1) In superior and anteroinferior plating of midclavicle fractures, which screw tips in a typical clavicular plating approach place the neurovascular structures at risk of injury? (2) How does patient positioning (supine or beach chair) affect the distance between the screws and the neurovascular structures? METHODS: The clavicles of 15 fresh-frozen cadavers were dissected. A hypothetical fracture line was marked at the midpoint of each clavicle. A precontoured six-hole 3.5-mm reconstruction locking compression plate was applied to the superior surface of the clavicle by using the fracture line to position the center of the plate. The direction of the drill bits and screws through screw holes that offer the greater risk of injury to the neurovascular structures were identified, and were defined as the risky screw holes, and the distances from the screw tips to the neurovascular structures were measured according to a standard protocol with a Vernier caliper in both supine and beach chair positions. Anteroinferior plating was also assessed following the same steps. The different distances from the screw tips to the neurovascular structures in the supine position were compared with the distances in the beach chair position using an unpaired t-test. RESULTS: The risky screw holes were the first medial and second medial screw holes. The relative distance ratios compared with the entire clavicular length for the distances from the sternoclavicular joint to the first medial and second medial screw holes were 0.46 and 0.36 in superior plating and 0.47 and 0.37 in anteroinferior plating, respectively. The riskiest screw hole for both superior and anteroinferior plates was the second medial screw hole in both the supine and beach chair positions (supine superior plating: 8.2 mm ± 3.1 mm [minimum: 1.1 mm]; beach chair anteroinferior plating: 7.6 mm ± 4.2 mm [minimum: 1.1 mm]). Patient positioning affected the distances between the riskiest screw tip and the nearest neurovascular structures, whereas in superior plating, changing from the supine position to the beach chair position increased this distance by 1.4 mm (95% CI -2.8 to -0.1; supine 8.2 ± 3.1 mm, beach chair 9.6 ± 2.1 mm; p = 0.037); by contrast, in anteroinferior plating, changing from the beach chair position to the supine position increased this distance by 5.4 mm (95% CI 3.6 to 7.4; beach chair 7.6 ± 4.2 mm, supine 13.0 ± 3.2 mm; p < 0.001). CONCLUSIONS: The second medial screw hole places the neurovascular structures at the most risk, particularly with superior plating in the supine position and anteroinferior plating in the beach chair position. CLINICAL RELEVANCE: The surgeon should be careful while making the first medial and second medial screw holes. Superior plating is safer to perform in the beach chair position, while anteroinferior plating is more safely performed in the supine position.

Proximity of the Coracoid Process to the Neurovascular Structures in Various Patient and Shoulder Positions: A Cadaveric Study.

PURPOSE: To examine and compare the distances from the anteromedial aspects of the coracoid base and the coracoid tip to the neurovascular structures in various patient positions. METHODS: The experiment was conducted in 15 fresh-frozen cadavers. We dissected 15 right and 15 left shoulders to measure the distances from the anteromedial aspects of the coracoid base and the coracoid tip to the lateral border of the neurovascular structures in the horizontal, vertical, and closest planes. The measurements were performed with the cadavers in the supine, lateral decubitus, and beach-chair positions. With cadavers in the beach-chair position, we evaluated 5 arm postures (arm at side, 45° of abduction, 90° of abduction, 45° of forward flexion, and 90° of forward flexion). RESULTS: The shortest distance from the coracoid base to the neurovascular structures was found in the beach-chair position with arm at side in the horizontal plane (27.4 ± 4.9 mm) and 90° of abduction in the vertical (21.8 ± 4.2 mm) and closest (19.5 ± 4.2 mm) planes. The distances in each plane were statistically significant compared with the supine and lateral decubitus positions (P < .005). Between the coracoid tip and the neurovascular structures, the shortest distance was found in the beach-chair position with 90° of abduction, with 29.3 ± 7.7 mm, 20.8 ± 4.9 mm, and 18.5 ± 5.1 mm in the horizontal, vertical, and closest planes, respectively. The distances were statistically significant in all planes compared with the supine and lateral decubitus positions (P < .005). CONCLUSIONS: Shoulder surgery in the area of the coracoid process is safe, especially with the patient in the supine position. The distance from the coracoid process to the neurovascular structures was closest in the beach-chair position with 90° of arm abduction. CLINICAL RELEVANCE: This study determined the distances between the coracoid process and the neurovascular structures during surgery around the coracoid process.

Prevention of Medial Femoral Condyle Injury by Using a Slotted Cannula in Anterior Cruciate Ligament Reconstruction.

Anterior cruciate ligament injury is one of the most common ligament injuries of the knee in sports medicine. The treatment of choice is anterior cruciate ligament reconstruction, which requires an experienced surgeon. Complications may occur at every step of the procedure. The surgeon should be especially careful when performing the step of femoral tunnel creation. Damage to the medial femoral condyle (MFC) could occur; thus, the full functional outcome of the knee might be affected. There are many surgical techniques to avoid MFC injury. We propose a surgical technique that could reduce the risk of MFC injury. This technique uses a Fast-Fix slotted cannula (Smith & Nephew) as a protector. Because such equipment is available in all meniscus instrument sets, this surgical technique is a feasible and easy method to use during the femoral tunnel creation step.