Search results for: bovine pericardium

Authors: Assiya Madenovna Borsynbayeva, Kairat Altynbekovich Turgenbayev, Nikolay Petrovich Ivanov

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In this article presents the results of rapid diagnostics of tuberculosis in comparison with classical bacteriological method. The proposed method of rapid diagnosis of tuberculosis than bacteriological method allows shortening the time of diagnosis to 7 days, to visualize the growth of mycobacteria in the semi-liquid medium and differentiate the type of mycobacterium. Fast definition of Mycobacterium tuberculosis and its derivatives in the culture medium is a new and promising direction in the diagnosis of tuberculosis.

Keywords: animal diagnosis of tuberculosis, bacteriological diagnostics, antigen, specific antibodies, immunological reaction

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Authors: Djeddi Khaled, Houssou Hind, Miloudi Abdellatif, Rabah Siham

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In the field of veterinary medicine, there is a growing application of artificial intelligence (AI) for diagnosing bovine mastitis, a prevalent inflammatory disease in dairy cattle. AI technologies, such as automated milking systems, have streamlined the assessment of key metrics crucial for managing cow health during milking and identifying prevalent diseases, including mastitis. These automated milking systems empower farmers to implement automatic mastitis detection by analyzing indicators like milk yield, electrical conductivity, fat, protein, lactose, blood content in the milk, and milk flow rate. Furthermore, reports highlight the integration of somatic cell count (SCC), thermal infrared thermography, and diverse systems utilizing statistical models and machine learning techniques, including artificial neural networks, to enhance the overall efficiency and accuracy of mastitis detection. According to a review of 15 publications, machine learning technology can predict the risk and detect mastitis in cattle with an accuracy ranging from 87.62% to 98.10% and sensitivity and specificity ranging from 84.62% to 99.4% and 81.25% to 98.8%, respectively. Additionally, machine learning algorithms and microarray meta-analysis are utilized to identify mastitis genes in dairy cattle, providing insights into the underlying functional modules of mastitis disease. Moreover, AI applications can assist in developing predictive models that anticipate the likelihood of mastitis outbreaks based on factors such as environmental conditions, herd management practices, and animal health history. This proactive approach supports farmers in implementing preventive measures and optimizing herd health. By harnessing the power of artificial intelligence, the diagnosis of bovine mastitis can be significantly improved, enabling more effective management strategies and ultimately enhancing the health and productivity of dairy cattle. The integration of artificial intelligence presents valuable opportunities for the precise and early detection of mastitis, providing substantial benefits to the dairy industry.

Keywords: artificial insemination, automatic milking system, cattle, machine learning, mastitis

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Authors: Balakrishna Valluru, Ruth Suckling

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A 16 year old male was found collapsed on the road following stab injuries to the chest and abdomen and was transported to the emergency department by ambulance. On arrival in the emergency department the patient was breathless and appeared pale. He was maintaining his airway with spontaneous breathing and had a heart rate of 122 beats per minute with a blood pressure of 83/63 mmHg. He was resuscitated initially with three units of packed red cells. Clinical examination identified three incisional wounds each measuring 2 cm. These were in the left para-sternal region, right infra-scapular region and left upper quadrant of the abdomen. The chest wound over the left parasternal area at the level of 4tth intercostal space was bleeding intermittently on leaning forwards and was relieving his breathlessness intermittently. CT imaging was performed to characterize his injuries and determine his management. CT scan of chest and abdomen showed moderate size haemopericardium with left sided haemopneumothorax. The patient underwent urgent surgical repair of the left ventricle and left anterior descending artery. He recovered without complications and was discharged from the hospital. This case highlights the fact that the potential to develop a life threatening cardiac tamponade was mitigated by the left parasternal stab wound. This injury fortuitously provided a pericardial window through which the bleeding from the injured left ventricle and left anterior descending artery could drain into the left hemithorax providing an opportunity for timely surgical intervention to repair the cardiac injuries.

Keywords: stab, incisional, haemo-pericardium, haemo-pneumothorax

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Authors: L. C. Prasanna, Antony Sylvan D’Souza, Kumar M. R. Bhat

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Although life as a conjoined twin would seem intolerable, there has recently been an increased interest in this subject because of the increasing number of cases where attempts have been made to separate them surgically. We have reviewed articles on cardiovascular anomalies in conjoined twins and presenting rarest anomaly in dicephalus parapagus fetus having two heads attached to one body from the neck or upper chest downwards, with a pair of limbs and a set of reproductive organs. Both the twins shared a common thoracic cavity with a single sternum. When the thoracic cavity was opened, a common anterior mediastinum was found. On opening the pericardium, two separate, closely apposed hearts were exposed. The two cardia are placed side by side. The left heart was slightly larger than the right and were joined at the atrial levels. Four atrial appendages were present, two for each twin. The atrial complex was a common chamber posterior to the ventricles. A single large tributary which could be taken as inferior vena cava drains into the common atrial chamber. In this case, the heart could not be assigned to either twin and therefore, it is referred to as the shared heart within a common pericardial sac. The right and left descending thoracic aorta have joined with each other just above the diaphragm to form a common descending thoracic aorta which has an opening in the diaphragm to be continued as common abdominal aorta which has a normal branching pattern. Upon an interior dissection, it is observed that the two atria have a wide communication which could be a wide patent foramen ovale and this common atrial cavity has a communication with a remnant of a possible common sinus venosus.

Keywords: atrium, congenital anomaly, conjoined twin, sinus venosus

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Authors: Ayeshmanthe Rathnayake, Siew Goh, Carmel Fenton, Ashutosh Hardikar

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Post-Operative Atrial Fibrillation (POAF) is the most frequent complication of cardiac surgery and is associated with reduced survival, increased rates of cognitive changes and cerebrovascular accident, heart failure, renal dysfunction, infection and length of stay, and hospital costs. Cardiac tamponade, although less common, carries high morbidity and mortality. Shed mediastinal blood in the pericardial space is a major source of intrapericardial oxidative stress and inflammation that triggers POAF. The utilisation of a left posterior pericardiotomy aims to shunt blood from the pericardium into the pleural space and have a role in the prevention of POAF as well as cardiac tamponade. 2118 patients had undergone isolated Coronary Artery Bypass Graft (CABG) at Royal Hobart Hospital from 2008-2021. They were divided into pericardiotomy vs control group. Patient baseline demographics, intraoperative data, and post-operative outcomes were reviewed retrospectively. Total incidence of new POAF and cardiac tamponade was 26.1% and 0.75%, respectively. Primary outcome of both the incidence of POAF(22.9% vs27.8%OR 0.77 p<0.05) and Cardiac Tamponade (0% vs 1.1% OR 0.85 p<0.05) were less in the pericardiotomy group.Increasing age, BMI, poor left ventricular function (EF <30%), and return to theatre were independent predictors of developing POAF. There were similar rates of return to theatre for bleeding however, no cases of tamponade in the pericardiotomy group. There were no complications attributable to left posterior pericardiotomy and the time added to the duration of surgery was minimal. Left posterior pericardiotomy is associated with a significant reduction in the incidence of POAFand cardiac tamponade and issafe and efficient.

Keywords: cardiac surgery, pericardiotomy, post-operative atrial fibrillation, cardiac tamponade

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Authors: Francisco J. Cedano, Laura M. Pinzón, Camila I. Castro, Felipe Salcedo, Juan P. Casas, Juan C. Briceño

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Complex fractures located in articular surfaces are challenging to treat and their reduction with conventional treatments could compromise the functionality of the affected limb. An adhesive material to treat those fractures is desirable for orthopedic surgeons. This adhesive must be biocompatible and have a high adhesion to bone surface in an aqueous environment. The proposed adhesive is based on chitosan, given its adhesive and biocompatibility properties. Chitosan is mixed with calcium carbonate and hydroxyapatite, which contribute to structural support and a gel like behavior, and glutaraldehyde is used as a cross-linking agent to keep the adhesive mechanical performance in aqueous environment. This work aims to evaluate the rheological, adhesion strength and biocompatibility properties of the proposed adhesive using in vitro tests. The gelification process of the adhesive was monitored by oscillatory rheometry in an ARG-2 TA Instruments rheometer, using a parallel plate geometry of 22 mm and a gap of 1 mm. Time sweep experiments were conducted at 1 Hz frequency, 1% strain and 37°C from 0 to 2400 s. Adhesion strength is measured using a butt joint test with bovine cancellous bone fragments as substrates. The test is conducted at 5 min, 20min and 24 hours after curing the adhesive under water at 37°C. Biocompatibility is evaluated by a cytotoxicity test in a fibroblast cell culture using MTT assay and SEM. Rheological results concluded that the average gelification time of the adhesive is 820±107 s, also it reaches storage modulus magnitudes up to 106 Pa; The adhesive show solid-like behavior. Butt joint test showed 28.6 ± 9.2 kPa of tensile bond strength for the adhesive cured for 24 hours. Also there was no significant difference in adhesion strength between 20 minutes and 24 hours. MTT showed 70 ± 23 % of active cells at sixth day of culture, this percentage is estimated respect to a positive control (only cells with culture medium and bovine serum). High vacuum SEM observation permitted to localize and study the morphology of fibroblasts presented in the adhesive. All captured fibroblasts presented in SEM typical flatted structure with filopodia growth attached to adhesive surface. This project reports an adhesive based on chitosan that is biocompatible due to high active cells presented in MTT test and these results were correlated using SEM. Also, it has adhesion properties in conditions that model the clinical application, and the adhesion strength do not decrease between 5 minutes and 24 hours.

Keywords: bioadhesive, bone adhesive, calcium carbonate, chitosan, hydroxyapatite, glutaraldehyde

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Authors: Rodrigo De A. Ferrazza, Henry D. M. Garcia, Viviana H. V. Aristizabal, Camilla De S. Nogueira, Cecilia J. Verissimo, Jose Roberto Sartori, Roberto Sartori, Joao Carlos P. Ferreira

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Environmental factors adversely influence sustainability in livestock production system. Dairy herds are the most affected by heat stress among livestock industries. This clearly implies in development of new strategies for mitigating heat, which should be based on physiological and metabolic adaptations of the animal. In this study, we incorporated the effect of climate variables and heat exposure time on the thermoregulatory responses in order to clarify the adaptive mechanisms for bovine heat dissipation under intense thermal stress induced experimentally in climate chamber. Non-lactating Holstein cows were contemporaneously and randomly assigned to thermoneutral (TN; n=12) or heat stress (HS; n=12) treatments during 16 days. Vaginal temperature (VT) was measured every 15 min with a microprocessor-controlled data logger (HOBO®, Onset Computer Corporation, Bourne, MA, USA) attached to a modified vaginal controlled internal drug release insert (Sincrogest®, Ourofino, Brazil). Rectal temperature (RT), respiratory rate (RR) and heart rate (HR) were measured twice a day (0700 and 1500h) and dry matter intake (DMI) was estimated daily. The ambient temperature and air relative humidity were 25.9±0.2°C and 73.0±0.8%, respectively for TN, and 36.3± 0.3°C and 60.9±0.9%, respectively for HS. Respiratory rate of HS cows increased immediately after exposure to heat and was higher (76.02±1.70bpm; P<0.001) than TN (39.70±0.71bpm), followed by rising of RT (39.87°C±0.07 for HS versus 38.56±0.03°C for TN; P<0.001) and VT (39.82±0.10°C for HS versus 38.26±0.03°C for TN; P<0.001). A diurnal pattern was detected, with higher (P<0.01) afternoon temperatures than morning and this effect was aggravated for HS cows. There was decrease (P<0.05) of HR for HS cows (62.13±0.99bpm) compared to TN (66.23±0.79bpm), but the magnitude of the differences was not the same over time. From the third day, there was a decrease of DMI for HS in attempt to maintain homeothermy, while TN cows increased DMI (8.27kg±0.33kg d-1 for HS versus 14.03±0.29kg d-1 for TN; P<0.001). By regression analysis, RT and RR better reflected the response of cows to changes in the Temperature Humidity Index and the effect of climate variables from the previous day to influence the physiological parameters and DMI was more important than the current day, with ambient temperature the most important factor. Comparison between acute (0 to 3 days) and chronic (13 to 16 days) exposure to heat stress showed decreasing of the slope of the regression equations for RR and DMI, suggesting an adaptive adjustment, however with no change for RT. In conclusion, intense heat stress exerted strong influence on the thermoregulatory mechanisms, but the acclimation process was only partial.

Keywords: acclimation, bovine, climate chamber, hyperthermia, thermoregulation

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Authors: Marcin Jakiel, Maria Kurek, Karolina Gutkowska, Sylwia Sanakiewicz, Dominika Stolarczyk, Jakub Batko, Rafał Jakiel, Mateusz K. Hołda

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The geometry of RVOT is extremely complicated. It is an irregular block with an ellipsoidal cross-section, whose dimensions decrease toward the pulmonary valve and measure 33.82 (IQR 30,51-39,36), 28.82 (IQR 26,11-32,22), 27.95 ± 4,11 for width [mm] and 33.41 ± 6,14, 26.99 ± 4,41, 26.91 ± 4,00 [mm] for depth, in the basal, middle and subpulmonary parts, respectively. In a sagittal section view, the RVOT heads upward and slightly backward. Its anterior perimeter has an average length of 41.96 mm and inclines to the transverse plane at an angle of 50.77° (IQR 46,53°-58,70°). In the posterior region, the RVOT is shorter (18.17mm) and flexes anteriorly. Therefore, the slope of the upper part of the rear wall to the transverse plane is an acute angle (open toward the rear) of 44,58° (IQR 37,30°-51,25°), while in the lower part it is an angle close to a right angle of 94,30°±15,44°. In addition, the thickness of the RVOT wall in the diastolic phase, at the posterior perimeter at the base, in the middle of the length and subpulmonary measure 3,80 mm ± 0,88 mm, 3,56 mm ± 0,73 mm, 3,56 mm ± 0,65 mm, respectively. In frontal cross-section, the RVOT rises on the interventricular septum, which makes it possible to distinguish the septal and supraseptal parts on its left periphery. The angles (facing the vertices to the right) of the inclination of these parts to the transverse plane are 75.5° (IQR 66,44°-81,11°) and 107.01° (IQR 99,09 – 115,23°), respectively, which allows us to conclude that the direction of the RVOT long axis changes from left to right. The above analysis shows that there is no single RVOT axis. Two axes can be distinguished, the one for the upper RVOT being more backward and leftward. The aforementioned forward deflection of the posterior wall and the RVOT's elevation over the interventricular septum, suggest that access to the subpulmonary region may be difficult. It should be emphasized that this area is often the target for ablation of ventricular arrhythmias. The small thickness of the RVOT posterior wall, with its difficult geometry, may favor its perforation into the pericardium or ascending aorta.

Keywords: angle, geometry, operation access, position, RVOT, shape

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Authors: Mohsen Shirani, Micaela Demichela

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During the last decades, a number of food crises such as Bovine Spongiform Encephalopathy (BSE), Mad-Cow disease, Dioxin in chicken food, Food-and-Mouth Disease (FMD), have certainly inflicted the reliability of the food industry. Consequently, the trend in applying different scientific methods of risk assessment in food safety has obtained more attentions in the academic and practice. However, lack of practical approach considering entire food supply chain is tangible in the academic literature. In this regard, this paper aims to apply risk assessment tool (FMEA) with integration of Human Factor along the entire supply chain of food production and test the method in a case study of Diary production, and analyze its results.

Keywords: FMEA, food supply chain, risk assessment, human factor

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Authors: Redie Kidane Ghebrehawariat, Betiel Habte Hadgu, Filmon Berhane Kahsay, Rim Berhane Fisehaye, Samuel Haile Kahsay, Saron Yemane Yosief, Selemawit Mosazghi Gilazghi

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A cross-sectional study was conducted from 22 February to 9 April 2022 on small, medium, and large holder dairy farms to determine the bovine mastitis prevalence and associated risk factors in the Anseba region, Eritrea. A total of 34 dairy farms and 193 dairy cows were randomly selected. Dairy cows were physically examined for any change on the udder and milk; a California mastitis test was performed to check sub-clinical mastitis; a closed-ended semi-structured questionnaire composed of 28 variables/risk factors (21 management risk factors and 7 animal-level risk factors) was used to determine the risk factors responsible for clinical and sub-clinical mastitis in the dairy cows. The overall cow-level prevalence of mastitis was 147 (76.2%). The animal level prevalence rate of clinical and sub-clinical mastitis was found to be 22 (11.4%) and 125 (64.8%), respectively, while herd level prevalence both for clinical and subclinical mastitis was found to be 14 (41.2%) and 26 (76.5%) respectively. Based on the already set P-value, which is <0.05, a number of risk factors were found to have a significant relationship with the occurrence of clinical and sub-clinical mastitis. Generally, animal risk factors such as animal age, parity, injury on the udder or teat, and previous history of mastitis presence of injury on the udder and lactation stage were risk factors with a significant relationship with the occurrence of clinical and sub-clinical mastitis. On the other hand, management risk factors with a significant relationship to the occurrence of clinical and sub-clinical mastitis were herd size, failure to milk mastitic cow, at last, educational level, floor type, failure to use a towel, using one towel for more than one cow and failure to practice mastitis test. From a total of 772 quarters, 280 (36.3%) were found positive for sub-clinical mastitis using the California mastitis test; of these, 70 (9%) were weakly positive, 90 (11.7%) were distinct positive, and 120 (15.5%) were strongly positive. Furthermore, 13 (1.7%) quarters were blocked. Quarter level prevalence was right front 80 (41.5%), left front 64 (33.3%), right hind 69 (35.8%) and left hind 67 (34.7%). The study has shown that mastitis is a major problem for dairy farms and the findings suggested that mastitis is one of the limiting factors in increasing milk production. Subclinical mastitis was found to be a devastating problem, and it occurred in all three breeds of lactating dairy cattle. Therefore, farmers should work hard to avoid the above-mentioned risk factors to minimize the infection of their dairy cattle by mastitis and thereby increase their profit. On the other hand, the Ministry of Agriculture, through the extension unit, should work in close contact with the farmers to increase awareness of the economic importance of the disease and associated risk factors.

Keywords: mastitis, prevalence, dairy cattle, Anseba, Eritrea

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Authors: Rensson Homero Celiz Ygnacio, Marco Aurélio Schiavo Novaes, Lucy Vanessa Sulca Ñaupas, Ana Paula Ribeiro Rodrigues

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The cryopreservation of testicular tissue emerges as an alternative for the preservation of the reproductive potential of individuals who still cannot produce sperm; however, they will undergo treatments that may affect their fertility (e.g., chemotherapy). Therefore, the present work aims to compare two cryopreservation methods (slow freezing and vitrification) in testicular tissue of prepubertal lambs. For that, to obtain the testicular tissue, the animals were castrated and the testicles were collected immediately in a physiological solution supplemented with antibiotics. In the laboratory, the testis was split into small pieces. The total size of the testicular fragments was 3×3x1 mm³ and was placed in a dish contained in Minimum Essential Medium (MEM-HEPES). The fragments were distributed randomly into non-cryopreserved (fresh control), slow freezing (SF), and vitrified. To SF procedures, two fragments from a given male were then placed in a 2,0 mL cryogenic vial containing 1,0 mL MEM-HEPES supplemented with 20% fetal bovine serum (FBS) and 20% dimethylsulfoxide (DMSO). Tubes were placed into a Mr. Frosty™ Freezing container with isopropyl alcohol and transferred to a -80°C freezer for overnight storage. On the next day, each tube was plunged into liquid nitrogen (NL). For vitrification, the ovarian tissue cryosystem (OTC) device was used. Testicular fragments were placed in the OTC device and exposed to the first vitrification solution composed of MEM-HEPES supplemented with 10 mg/mL Bovine Serum Albumin (BSA), 0.25 M sucrose, 10% Ethylene glycol (EG), 10% DMSO and 150 μM alpha-lipoic acid for four min. The VS1 was discarded and then the fragments were submerged into a second vitrification solution (VS2) containing the same composition of VS1 but 20% EG and 20% DMSO. VS2 was then discarded and each OTC device containing up to four testicular fragments was closed and immersed in NL. After the storage period, the fragments were removed from the NL, kept at room temperature for one min and then immersed at 37 °C in a water bath for 30 s. Samples were warmed by sequentially immersing in solutions of MEM-HEPES supplemented with 3 mg/mL BSA and decreasing concentrations of sucrose. Hematoxylin-eosin staining to analyze the tissue architecture was used. The score scale used was from 0 to 3, classified with a score 0 representing normal morphologically, and 3 were considered a lot of alteration. The histomorphological evaluation of the testicular tissue shows that when evaluating the nuclear alteration (distinction of nucleoli and condensation of nuclei), there are no differences when using slow freezing with respect to the control. However, vitrification presents greater damage (p <0.05). On the other hand, when evaluating the epithelial alteration, we observed that the freezing showed scores statistically equal to the control in variables such as retraction of the basement membrane, formation of gaps and organization of the peritubular cells. The results of the study demonstrated that cryopreservation using the slow freezing method is an excellent tool for the preservation of pubertal testicular tissue.

Keywords: cryopreservation, slow freezing, vitrification, testicular tissue, lambs

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Authors: Mohan Singh Mehata, R. K. Ratnesh

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CdSe, CdSe/ZnS, and CdSe/CdS core-shell quantum dots (QDs) of 3-4 nm were developed by using chemical route and following successive ion layer adsorption and reaction (SILAR) methods. The prepared QDs have been examined by using X-ray diffraction, high-resolution electron microscopy and optical spectroscopy. The photoluminescence (PL) quantum yield (QY) of core-shell QDs increases with respect to the core, indicating that the radiative rate increases by the formation of shell around core, as evident by the measurement of PL lifetime. Further, the PL of bovine serum albumin is quenched strongly by the presence of core-shall QDs and follow the Stern-Volmer (S-V) relation, whereas the lifetime does not follow the S-V relation, demonstrating that the observed quenching is predominantly static in nature. Among all the QDs, the CdSe/ZnS QDs shows the least cytotoxicity hence most biocompatibility.

Keywords: biocompatibility, core-shell quantum dots, photoluminescence and lifetime, sensing ability

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Authors: M. Akoz, O. B. Citil, I. Aydin

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Colostrum secreted by the mammary glands after birth in the early days, a high proportion of fat, protein and ash containing a secretion containing low amounts of casein and lactose. Especially immunoglobulins contain high proportions. Maternal immunoglobulins own immune system to protect the newborn against neonatal disease until development are very important matter. However, colostrum is transferred to the offspring due to placental barrier in ruminants. Immunoglobulins are absorbed through the intestinal epithelium but absorption can vary under the influence of some factors. These factors are among the priority ones taking colostrum first time, amount, concentration, the metabolic status of the newborn. intestinal absorption of immunoglobulins occurs over the first 24 h high. Absorption from the gut after nine hours, 50% after 24 hours was only 11%. On the other hand pup's digestive system degrade the enzymes after 24 hours immunoglobulins. Bovine colostrum in the composition while basic immune IgG, IgA and IgM are also available. Total IgG in colostrum of ruminants, while in other species is a greater amount in blood serum.

Keywords: immunoglobulin, ruminants, colostrum, immune system

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Authors: Agnė Mikalauskaitė, Renata Karpicz, Vitalijus Karabanovas, Arūnas Jagminas

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The use of biocompatible precursors for the synthesis and stabilization of fluorescent gold nanoclusters (NCs) with strong red photoluminescence creates an important link between natural sciences and nanotechnology. Herein, we report the cost-effective synthesis of Au nanoclusters by templating and reduction of chloroauric acid with the cheap amino acid food supplements. This synthesis under the optimized conditions leads to the formation of biocompatible Au NCs having good stability and intense red photoluminescence, peaked at 680 to 705 nm, with a quantum yield (QY) of ≈7% and the average lifetime of up to several µs. The composition and luminescent properties of the obtained NCs were compared with ones formed via well-known bovine serum albumin reduction approach. Our findings implied that synthesized Au NCs tend to accumulate in more tumorigenic breast cancer cells (line MDA-MB-213) and after dialysis can be prospective for bio imagining.

Keywords: gold nanoclusters, proteins, materials chemistry, red-photoluminescence, bioimaging

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Authors: Chih-Chi Cheng, Ji-Yu Lin, I-Ming Chu

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In this study, we synthesized a thermosensitive polypeptide hydrogel by copolymerizing poloxamer (PLX) and poly(ʟ-alanine) with ʟ-lysine segments at the both ends to form PLX-b-poly(ʟ-alanine-lysine) (Lys-Ala-PLX-Ala-Lys) copolymers. Poly(ʟ-alanine) is the hydrophobic chain of Lys-Ala-PLX-Ala-Lys copolymers which was designed to capture the hydrophobic agents. The synthesis was examined by 1H NMR and showed that Lys-Ala-PLX-Ala-Lys copolymers were successfully synthesized. At the concentration range of 3-7 wt%, the aqueous copolymer solution underwent sol-gel transition near the physiological temperature and exhibited changes in its secondary structure content, as evidenced by FTIR. The excellent viability of cells cultured within the scaffold was observed after 72 hr of incubation. Also, negatively charged bovine serum albumin was incorporated into the hydrogel without diminishing material integrity and shows good release profile. In the animal study, the results also indicated that Lys-Ala-PLX-Ala-Lys hydrogel has high potential in wound dressing.

Keywords: polypeptide thermosensitive hydrogel, tacrolimus, vascularized composite allotransplantation, sustain release

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Authors: Vasile Simulescu, Jakub Mondek, Miloslav Pekař

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Hyaluronic acid (HA) is an anionic glycosaminoglycan distributed throughout connective, epithelial and neural tissues. The importance of hyaluronic acid increased in the last decades. It has many applications in medicine and cosmetics. Hyaluronic acid has been used in attempts to treat osteoarthritis of the knee via injecting it into the joint. Bovine serum albumin (also known as BSA) is a protein derived from cows, which has many biochemical applications. The aim of our research work was to compare the thermal degradation of hyaluronic acid and BSA in powder and in solution, by determining changes in molar mass and conformation, by using SEC-MALLS (size exclusion chromatography -multi angle laser light scattering). The aim of our research work was to observe the degradation in powder and in solution of different molar mass hyaluronic acid samples, at different temperatures for certain periods. The degradation of the analyzed samples was mainly observed by modifications in molar mass.

Keywords: thermal degradation, hyaluronic acid, BSA, SEC-MALLS

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Authors: Loredana E. Nita, Aurica P. Chiriac, Elena Stoleru, Alina Diaconu, Tudorachi Nita

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Self-assembly processes are an attractive method to form new and complex structures between macromolecular compounds to be used for specific applications. In this context, intramolecular and intermolecular bonds play a key role during self-assembling processes in preparation of carrier systems of bioactive substances. Polyelectrolyte complexes (PECs) are formed through electrostatic interactions, and though they are significantly below of the covalent linkages in their strength, these complexes are sufficiently stable owing to the association processes. The relative ease way of PECs formation makes from them a versatile tool for preparation of various materials, with properties that can be tuned by adjusting several parameters, such as the chemical composition and structure of polyelectrolytes, pH and ionic strength of solutions, temperature and post-treatment procedures. For example, protein-polyelectrolyte complexes (PPCs) are playing an important role in various chemical and biological processes, such as protein separation, enzyme stabilization and polymer drug delivery systems. The present investigation is focused on evaluation of the PPC formation between a synthetic polypeptide (poly(aspartic acid) – PAS) and a natural protein (bovine serum albumin - BSA). The PPC obtained from PAS and BSA in different ratio was investigated by corroboration of various techniques of characterization as: spectroscopy, microscopy, thermo-gravimetric analysis, DLS and zeta potential determination, measurements which were performed in static and/or dynamic conditions. The static contact angle of the sample films was also determined in order to evaluate the changes brought upon surface free energy of the prepared PPCs in interdependence with the complexes composition. The evolution of hydrodynamic diameter and zeta potential of the PPC, recorded in situ, confirm changes of both co-partners conformation, a 1/1 ratio between protein and polyelectrolyte being benefit for the preparation of a stable PPC. Also, the study evidenced the dependence of PPC formation on the temperature of preparation. Thus, at low temperatures the PPC is formed with compact structure, small dimension and hydrodynamic diameter, close to those of BSA. The behavior at thermal treatment of the prepared PPCs is in agreement with the composition of the complexes. From the contact angle determination results the increase of the PPC films cohesion, which is higher than that of BSA films. Also, a higher hydrophobicity corresponds to the new PPC films denoting a good adhesion of the red blood cells onto the surface of PSA/BSA interpenetrated systems. The SEM investigation evidenced as well the specific internal structure of PPC concretized in phases with different size and shape in interdependence with the interpolymer mixture composition.

Keywords: polyelectrolyte – protein complex, bovine serum albumin, poly(aspartic acid), self-assembly

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Authors: Munirah Ismail, Joon Hock Yeo

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Paediatric patients who require ventricular outflow tract reconstruction usually need valve construction to prevent valvular regurgitation. They would face problems like lack of suitable, affordable conduits and the need to undergo several operations in their lifetime due to the short lifespan of existing valves. Their natural growth and development are also of concern, even if they manage to receive suitable conduits. Current prosthesis including homografts, bioprosthetic valves, mechanical valves, and bovine jugular veins either do not have the long-term durability or the ability to adapt to the growth of such patients. We have developed a new design of bi-leaflet valve. This new technique accommodates patients’ annular size growth while maintaining valvular patency. A mock circulatory system was set up to assess the hemodynamic performance of the bi-leaflet pulmonary valve. It was found that the percentage regurgitation was acceptable and thus, validates this novel concept.

Keywords: bi-leaflet pulmonary valve, pulmonary heart valve, tetralogy of fallot, mock circulatory system

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Authors: Djamel Remache, Marie Semaan, Cécile Baron, Martine Pithioux, Patrick Chabrand, Jean-Marie Rossi, Jean-Louis Milan

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Cortical bone is a complex multi-scale structure. Even though several works have contributed significantly to understanding its mechanical behavior, this behavior remains poorly understood. Nanoindentation testing is one of the primary testing techniques for the mechanical characterization of bone at small scales. The purpose of this study was to provide new nanoindentation data of cortical bovine bone in different directions and at different bone microstructures (osteonal, interstitial and laminar bone), and then to identify anisotropic properties of samples with FEM (finite element method) based inverse method. Experimentally and numerical results were compared. Experimental and numerical results were compared. The results compared were in good agreement.

Keywords: mechanical behavior of bone, nanoindentation, finite element analysis, inverse optimization approach

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Authors: Hamid Mustafa, Heather J. Huson, Adeela Ajmal, Kim Euisoo, Tad S. Sonstegard

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In this study, 67 animals, representing six different cattle breeds of Pakistan, were genotyped with the Bovine high density (777K) SNP Beadchip. These include 13 Sahiwal, 09 Red Sindhi, 13 Tharparkar, 08 Achi, 13 Cholistani and 10 Dhanni cattle breeds. Analysis of 500, 939 SNP markers revealed that the mean minor allele frequency (MAF) was 0.21, 0.22, 0.18, 0.23, 0.22 and 0.22 for Sahiwal, Red Sindhi, Tharparkar, Achi, Cholistani and Dhanni respectively. Significant differences of minor allele frequency (MAF) were observed between the indigenous Pakistani cattle population (P<0.001). Across these Pakistani cattle breeds, a common variant MAF (≥0.10 and ≤0.5) accounted for an overall estimated 75.71 % of the 500,939 SNPs and on the average 19.58 % of the markers were monomorphic. Mean observed (HO) and expected (HE) heterozygosities were 0.656 and 0.638, respectively. This primarily study of Pakistani indigenous cattle breeds indicate that this level of SNPs variation can potentially be used for genomic studies for future breeding plans and for farm animal conservation strategies.

Keywords: Pakistan, cattle, minor allele frequency, SNP, variation

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Authors: Alena Semeradtova, Marcel Stofik, Lucie Mareckova, Petr Maly, Ondrej Stanek, Jan Maly

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Recently, novel strategies based on so-called molecular evolution were shown to be effective for the production of various peptide ligand libraries with high affinities to molecular targets of interest comparable or even better than monoclonal antibodies. The major advantage of these peptide scaffolds is mainly their prevailing low molecular weight and simple structure. This study describes a new high-affinity binding molecules based immunesensor using a simple optical system for human serum albumin (HSA) detection as a model molecule. We present a comparison of two variants of recombinant binders based on albumin binding domain of the protein G (ABD) performed on micropatterned glass chip. Binding domains may be tailored to any specific target of interest by molecular evolution. Micropatterened glass chips were prepared using UV-photolithography on chromium sputtered glasses. Glass surface was modified by (3-aminopropyl)trietoxysilane and biotin-PEG-acid using EDC/NHS chemistry. Two variants of high-affinity binding molecules were used to detect target molecule. Firstly, a variant is based on ABD domain fused with TolA chain. This molecule is in vivo biotinylated and each molecule contains one molecule of biotin and one ABD domain. Secondly, the variant is ABD domain based on streptavidin molecule and contains four gaps for biotin and four ABD domains. These high-affinity molecules were immobilized to the chip surface via biotin-streptavidin chemistry. To eliminate nonspecific binding 1% bovine serum albumin (BSA) or 6% fetal bovine serum (FBS) were used in every step. For both variants range of measured concentrations of fluorescently labelled HSA was 0 – 30 µg/ml. As a control, we performed a simultaneous assay without high-affinity binding molecules. Fluorescent signal was measured using inverse fluorescent microscope Olympus IX 70 with COOL LED pE 4000 as a light source, related filters, and camera Retiga 2000R as a detector. The fluorescent signal from non-modified areas was substracted from the signal of the fluorescent areas. Results were presented in graphs showing the dependence of measured grayscale value on the log-scale of HSA concentration. For the TolA variant the limit of detection (LOD) of the optical immunosensor proposed in this study is calculated to be 0,20 µg/ml for HSA detection in 1% BSA and 0,24 µg/ml in 6% FBS. In the case of streptavidin-based molecule, it was 0,04 µg/ml and 0,07 µg/ml respectively. The dynamical range of the immunosensor was possible to estimate just in the case of TolA variant and it was calculated to be 0,49 – 3,75 µg/ml and 0,73-1,88 µg/ml respectively. In the case of the streptavidin-based the variant we didn´t reach the surface saturation even with the 480 ug/ml concentration and the upper value of dynamical range was not estimated. Lower value was calculated to be 0,14 µg/ml and 0,17 µg/ml respectively. Based on the obtained results, it´s clear that both variants are useful for creating the bio-recognizing layer on immunosensors. For this particular system, it is obvious that the variant based on streptavidin molecule is more useful for biosensing on glass planar surfaces. Immunosensors based on this variant would exhibit better limit of detection and wide dynamical range.

Keywords: high affinity binding molecules, human serum albumin, optical immunosensor, protein G, UV-photolitography

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Authors: Lizeth Fuentes-Mera, Vanessa Perez-Silos, Nidia K. Moncada-Saucedo, Alejandro Garcia-Ruiz, Alberto Camacho, Jorge Lara-Arias, Ivan Marino-Martinez, Victor Romero-Diaz, Adolfo Soto-Dominguez, Humberto Rodriguez-Rocha, Hang Lin, Victor Pena-Martinez

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Biphasic scaffolds in cartilage tissue engineering have been designed to influence not only the recapitulation of the osteochondral architecture but also to take advantage of the healing ability of bone to promote the implant integration with the surrounding tissue and then bone restoration and cartilage regeneration. This study reports the development and characterization of a biphasic scaffold based on the assembly of a cartilage phase constituted by fibroin biofunctionalized with bovine cartilage matrix; cellularized with differentiated pre-chondrocytes from adipose tissue stem cells (autologous) and well attached to a bone phase (bone bovine decellularized) to mimic the structure of the nature of native tissue and to promote the cartilage regeneration in a model of joint damage in pigs. Biphasic scaffolds were assembled by fibroin crystallization with methanol. The histological and ultrastructural architectures were evaluated by optical and scanning electron microscopy respectively. Mechanical tests were conducted to evaluate Young's modulus of the implant. For the biological evaluation, pre-chondrocytes were loaded onto the scaffolds and cellular adhesion, proliferation, and gene expression analysis of cartilage extracellular matrix components was performed. The scaffolds that were cellularized and matured for 10 days were implanted into critical 3 mm in diameter and 9-mm in depth osteochondral defects in a porcine model (n=4). Three treatments were applied per knee: Group 1: monophasic cellular scaffold (MS) (single chondral phase), group 2: biphasic scaffold, cellularized only in the chondral phase (BS1), group 3: BS cellularized in both bone and chondral phases (BS2). Simultaneously, a control without treatment was evaluated. After 4 weeks of surgery, integration and regeneration tissues were analyzed by x-rays, histology and immunohistochemistry evaluation. The mechanical assessment showed that the acellular biphasic composites exhibited Young's modulus of 805.01 kPa similar to native cartilage (400-800 kPa). In vitro biological studies revealed the chondroinductive ability of the biphasic implant, evidenced by an increase in sulfated glycosaminoglycan (GAGs) and type II collagen, both secreted by the chondrocytes cultured on the scaffold during 28 days. No evidence of adverse or inflammatory reactions was observed in the in vivo trial; however, In group 1, the defects were not reconstructed. In group 2 and 3 a good integration of the implant with the surrounding tissue was observed. Defects in group 2 were fulfilled by hyaline cartilage and normal bone. Group 3 defects showed fibrous repair tissue. In conclusion; our findings demonstrated the efficacy of biphasic and bioactive scaffold based on silk fibroin, which entwined chondroinductive features and biomechanical capability with appropriate integration with the surrounding tissue, representing a promising alternative for osteochondral tissue-engineering applications.

Keywords: biphasic scaffold, extracellular cartilage matrix, silk fibroin, osteochondral tissue engineering

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Authors: C. I. Boshoff, S. Steenkamp-Jonker

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Cystic echinococcosis (CE), caused by Echinococcus granulosus is an important parasitic infection in livestock worldwide, with severe zoonotic potential. It is important to understand the variability of Echinococcus granulosus, as genotype variations may influence lifecycle patterns, development rate, and transmission. Cystic Echinococcus samples were collected from domestic animals in Eastern Cape Province, South Africa. A molecular study was performed on 14 hydatid cysts obtained from caprine, ovine and bovine livers in order to determine the Echinococcus granulosus strain present in these hosts. The sequencing of the mitochondrial cytochrome C oxidase subunit I (coxI) gene of the hydatid cysts produced sequences of 400 bp for each sample analysed. These sequences were aligned with those present in GenBank and a phylogenetic tree was constructed. Based on coxI genotype the isolates could be grouped into E. granulosus sensu stricto. The findings of the study represent a pilot molecular study on Echinococcus from domestic animals undertaken in South Africa.

Keywords: Echinococcus granulosus, genotypes, livestock, South Africa

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Authors: Binnur Sagbas

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Artificial joint replacements such as total knee and total hip prosthesis have been applied to the patients who affected by osteoarthritis. Although different material combinations are used for these joints, biopolymers are most commonly preferred materials especially for acetabular cup and tibial component of hip and knee joints respectively. The main limitation that shortens the service life of these prostheses is wear. Wear is complicated phenomena and it must be considered with friction and lubrication. In this study, micro wave (MW) induced argon+oxygen plasma surface modification were applied on ultra-high molecular weight polyethylene (UHMWPE) and vitamin E blended UHMWPE (VE-UHMWPE) biopolymer surfaces to improve surface wettability and wear resistance of the surfaces. Contact angel measurement method was used for determination of wettability. Ball-on-disc wear test was applied under 25% bovine serum lubrication conditions. The results show that surface wettability and wear resistance of both material samples were increased by plasma surface modification.

Keywords: artificial joints, plasma surface modification, UHMWPE, vitamin E, wear

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Authors: Mosaab A. Omar, Mohammad Saleh Al-Aboody, Mohmed A. Rizk, Shimaa M. Elsayed, Ahmed ElSify, Naoaki Yokoyama, Ikuo Igarashi

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Enoxacin is a broad-spectrum 6-fluoronaphthyridinone antibacterial agent (fluoroquinolones) structurally related to nalidixic acid used mainly in the treatment of urinary tract infections and gonorrhea. Also, it has been shown recently that it may have cancer inhibiting effect. The primary antibabesial effect of Enoxacin is due to inhibition of DNA gyrase subunit A, and DNA topoisomerase. In the present study, enoxacin was tested as a potent inhibitor against the in vitro growth of bovine and equine Piroplasms. The in vitro growth of five Babesia species that were tested was significantly inhibited (P<0.05) by micromolar concentrations of enoxacin (IC50 values= 13.5, 7.2, 7.5, and 24.2 µM for Babesia bovis, Babesia bigemina, Babesia caballi, and Theileria equi, respectively). Enoxacin IC50 values for Babesia and Theileria parasites were satisfactory as the drug is a potent antibacterial drug with minimum side effects. Therefore, enoxacin might be used for the treatment of Babesiosis and Theileriosis especially in case of mixed infections with bacterial diseases or in the case of animal sensitivity against diminazin toxicity.

Keywords: enoxacin, Babesia, Theileria, IC50 and dimenazin

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Authors: Rima Zakzouk, Noriko Hiraishi, Mohamed Mahdi Alshahni, Koichi Makimura, Junji Tagami

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Background and Purpose: Silver diamine fluoride (SDF) is used to prevent and arrest dental caries. The aim of this study is to evaluate the effect of SDF on reducing Candida albicans adhesion on dentin. Materials and Methods: Bovine dentin disks (6×6 mm) were cut by Isomet and polished using grit silicon carbide papers down to 2000 in order to obtain flat dentin surfaces. Samples were divided into two groups. The first group (SDF group) was treated with 38% SDF for 3 min, while the other group (control group) did not undergo SDF treatment. All samples were exposed to C. albicans suspension, washed after 6 hours incubation at 30 °C before to be tested using XTT (2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide) and real time PCR approaches. Statistical analyses of the results were performed at the significance level α = 0.05. Results: SDF inhibited C. albicans adhesion onto dentin. A significant difference was found between the SDF and control groups in both XTT and real time PCR tests. Conclusion: Using SDF to arrest the caries, could inhibit the Candida growth on dentin.

Keywords: silver diamine fluoride, dentin, real time PCR, XTT

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Authors: Mohammad Asadpour, Gholamreza Razmi, Gholamreza Mohammadi, Abolghasem Naghibi

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Cryptosporidium Spp., protozoan parasites of the phylum Apicomplexa, have a wide spectrum of hosts including humans, domestic animals and wild mammals, birds, reptiles, amphibians and fish. Dairy cattle have been identified in numerous reports as a major source of environmental contamination with this pathogen. In this study, a Polymerase Chain Reaction (PCR), Restriction Fragment Length Polymorphism (RFLP) analysis of the Small-Subunit (SSU) rRNA gene was used to detect and identify Cryptosporidium Spp. in 300 fecal specimens from 1 to 30 days pre-wean calves in 10 farms in Mashhad, Iran. Eighty five (28.3%) and forty five (15%) of the specimens were positive for Cryptosporidium by microscopic and PCR examination respectively. Restriction digestion of the PCR products by VSPI and Ssp1 restriction enzymes and analysis of sequence data revealed the presence of C. parvum, bovine genotype in all isolates. Our findings suggest that cattle can be a source of Cryptosporidial infections for humans and animals in Mashhad area. This is the first published description of Cryptosporidium sub genotyping in Mashhad.

Keywords: cryptosporidium, genotype, dairy calves, 18S rRNA, Mashhad

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Authors: S. Md. Shaarani, J. Md. Jahim, R. A. Rahman, R. Md. Illias

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Modern developments in biotechnology have paved the way for extensive use of biocatalysis in industries. Although it offers immense potential, industrial application is usually hampered by lack of operational stability, difficulty in recovery as well as limited re-use of the enzyme. These drawbacks, however, can be overcome by immobilization. Cross-linked enzyme aggregates (CLEAs), a versatile carrier-free immobilization technique is one that is currently capturing global interest. This approach involves precipitating soluble enzyme with an appropriate precipitant and subsequent crosslinking by a crosslinking reagent. Without ineffective carriers, CLEAs offer high enzymatic activity, stability and reduced production cost. This study demonstrated successful CLEA synthesis of recombinant xylanase from Trichoderma reesei using ethanol as aggregating agent and glutaraldehyde (2% (v/v); 100 mM) as crosslinker. Effects of additives including proteic feeder such as bovine serum albumin (BSA) and poly-L-Lysine were investigated to reveal its significance in enhancing the performance of enzyme. Addition of 0.1 mg BSA/U xylanase showed considerable increment in CLEA development with approximately 50% retained activity.

Keywords: cross-linked, immobilization, recombinant, xylanase

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Authors: Jessica P. Fiorotti, Maria C. Freitas, Caio J. B. Coutinho-Rodrigues, Mariana G. Camargo, Emily S. Mesquita, Amanda R. C. Corval, Ricardo O. B. Bitencourt, Allan F. Marciano, Diva D. Spadacci-Morena, Patricia S. Golo, Isabele C. Angelo, Vania R. E. P. Bittencourt

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The bovine tick, Rhipicephalus microplus, is an arthropod of great importance in veterinary medicine leading to anemia, weight loss, animals' leather depreciation and also acting as a vector of many pathogens. In this way, the parasitism causes a loss of 3.24 billion dollars per year in Brazil. Knowingly, entomopathogenic fungi act as natural controller of some arthropods, acting mainly by active penetration through the cuticle. However, it can also act on the hemolymph and through the production of mycotoxins. Hemocytes are responsible for the cellular immune response and participate in the processes of phagocytosis, nodulation and encapsulation and may undergo changes when challenged by pathogens. The aim of the present study was to evaluate changes in R. microplus hemocytes after inoculation of Metarhizium robertsii using transmission electron microscopy. The isolate ARSEF 2575 and 200 engorged R. microplus females were used. The groups were divided into control, in which the females were inoculated with 5 μL of sterile distilled water solution and 0.1% Tween 80, and a group inoculated with 5 μL of fungal suspension at the concentration of 10⁷ conidia mL⁻¹. The experiment was performed in duplicate and each group contained 50 females. Twenty-four hours after fungal inoculation, hemolymph was collected through the cuticle dorsal surface perforation of the tick females. After collection, the hemolymph samples were centrifuged at 500 x g for 3 minutes at 4 °C, the plasma was discarded and the hemocyte pellet was resuspended in 50 μl PBS. The suspension material was fixed in 2% glutaraldehyde in Millonig buffer for three hours. After fixation, the material was centrifuged at 500 x g for 3 minutes, the supernatant was discarded and the cells were resuspended in a wash solution. Subsequently, the cells were post-fixed with 1% osmium tetroxide in phosphate buffer for one hour at room temperature and dehydrated in increasing concentrations of ethanol, and then embedded in Epon resin. The ultrathin sections were examined under the LEO EM 906E transmission electron microscopy at 80kV. The ultrastructural results revealed that.in control group, the cells were considered intact, in which the granulocytes were observed with granules of different electrodensities, intact mitochondria and cytoplasm without vacuolization. In addition, granulocytes showed plasma membrane projections similar to pseudopodia. Plasmatocytes presented as irregularly shaped cells, with the eccentric nucleus, agranular cytoplasm and some cells presented pseudopodia. Nevertheless, in the group exposed to the fungus, most of the cells presented in degeneration. The granulocytes found had fewer granules in the cytoplasm and more vacuoles. Plasmatocytes, after treatment, presented many vacuoles also in the cytoplasm and the lysosomes presented great amount of electrodense material in their interior. Thus, the results suggest that the fungus has a depressant action in the immune system of the tick, not only by the cell degranulation, but also suggesting that this leads to morphological changes in the hemocytes and may even trigger processes such as phagocytosis.

Keywords: bovine tick, cellular defense, entomopathogenic fungi, immune response

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Authors: J. Suwanprateeb, F. Thammarakcharoen

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Calcium phosphate coating (CaP) has been employed for protein delivery, but the typical direct protein adsorption on the coating led to low incorporation content and fast release of the protein from the coating. By using bovine serum albumin (BSA) as a model protein, rapid biomimetic co-precipitation between calcium phosphate and BSA was employed to control the distribution of BSA within calcium phosphate coating during biomimetic formation on titanium surface for only 6 h at 50 oC in an accelerated calcium phosphate solution. As a result, the amount of BSA incorporation and release duration could be increased by using a rapid biomimetic co-precipitation technique. Up to 43 fold increases in the BSA incorporation content and the increase from 6 h to more than 360 h in release duration compared to typical direct adsorption technique were observed depending on the initial BSA concentration used during co-precipitation (1, 10, and 100 microgram/ml). From X-ray diffraction and Fourier transform infrared spectroscopy studies, the coating composition was not altered with the incorporation of BSA by this rapid biomimetic co-precipitation and mainly comprised octacalcium phosphate and hydroxyapatite. However, the microstructure of calcium phosphate crystals changed from straight, plate-like units to curved, plate-like units with increasing BSA content.

Keywords: biomimetic, Calcium Phosphate Coating, protein, titanium

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