Search results for: bovine hemoglobin

Authors: Seong Gu Hwang, Young Kyoon Oh, Joseph F. dela Cruz

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Marbling, or intramuscular fat, has been consistently identified as one of the top beef quality problems. Intramuscular adipocytes distribute throughout the perimysial connective tissue of skeletal muscle and are the major site for the deposition of intramuscular fat, which is essential for the eating quality of meat. The stromal vascular fraction of the skeletal muscle contains progenitor cells that can be enhanced to differentiate to adipocytes and increase intramuscular fat. Primary cultures of bovine intramuscular stromal vascular cells were used in this study to elucidate the effects of extracellular calcium and retinoic acid concentration on adipocyte differentiation. Cell viability assay revealed that even at different concentrations of calcium and retinoic acid, there was no significant difference on cell viability. Monitoring of the adipocyte differentiation showed that bovine intramuscular stromal vascular cells cultured in a low concentration of extracellular calcium and retinoic acid had a better degree of fat accumulation. The mRNA and protein expressions of PPARγ, C/EBPα, SREBP-1c and aP2 were analyzed and showed a significant upregulation upon the reduction in the level of extracellular calcium and retinoic acid. The upregulation of these adipogenic related genes means that the decreasing concentration of calcium and retinoic acid is able to stimulate the adipogenic differentiation of bovine intramuscular stromal vascular cells. To further elucidate the effect of calcium, the expression level of calreticulin was measured. Calreticulin which is known to be an inhibitor of PPARγ was down regulated by the decreased level of calcium and retinoic acid in the culture media. The same tendency was observed on retinoic acid receptors RARα and CRABP-II. These receptors are recognized as adipogenic inhibitors, and the downregulation of their expression allowed a better level of differentiation in bovine intramuscular stromal vascular cells. In conclusion, data show that decreasing the level of extracellular calcium and retinoic acid can significantly promote adipogenesis in intramuscular stromal vascular cells of Hanwoo beef cattle. These findings may provide new insights in enhancing intramuscular adipogenesis and marbling in beef cattle.

Keywords: calcium, calreticulin, hanwoo beef, retinoic acid

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Authors: Haiyoung Jung, Mi Joung Leem, Sun Hwa Lee

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Background & Objective: Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a genetic abnormality that results in an inadequate amount of G6PD, leading to increased susceptibility of red blood cells to reactive oxygen species and hemolysis. The present study aimed to evaluate the careSTARTTM S1 analyzer for measuring G6PD activity to hemoglobin (Hb) ratio. Methods: Precision for G6PD activity and hemoglobin measurement was evaluated using control materials with two levels on five repeated runs per day for five days. The analytic performance of the careSTARTTM S1 analyzer was compared with spectrophotometry in 40 patient samples. Reference ranges suggested by the manufacturer were validated in 20 healthy males and females each. Results: The careSTARTTM S1 analyzer demonstrated precision of 6.0% for low-level (14~45 U/dL) and 2.7% for high-level (60~90 U/dL) control in G6PD activity, and 1.4% in hemoglobin (7.9~16.3 u/g Hb). A comparison study of G6PD to Hb ratio between the careSTARTTM S1 analyzer and spectrophotometry showed an average difference of 29.1% with a positive bias of the careSTARTTM S1 analyzer. All normal samples from the healthy population were validated for the suggested reference range for males (≥2.19 U/g Hb) and females (≥5.83 U/g Hb). Conclusion: The careSTARTTM S1 analyzer demonstrated good analytical performance and can replace the current spectrophotometric measurement of G6PD enzyme activity. In the aspect of the management of clinical laboratories, it can be a reasonable option as a point-of-care analyzer with minimal handling of samples and reagents, in addition to the automatic calculation of the ratio of measured G6PD activity and Hb concentration, to minimize any clerical errors involved with manual calculation.

Keywords: POCT, G6PD, performance evaluation, careSTART

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Authors: Dagmara A. Niedziela, Mark P. Murphy, Orla M. Keane, Finola C. Leonard

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Staphylococcus aureus is the most frequent cause of clinical and subclinical bovine mastitis in Ireland. Mastitis caused by S. aureus is often chronic and tends to recur after antibiotic treatment. This may be due to several virulence factors, including attributes that enable the bacterium to internalize into bovine mammary epithelial cells, where it may evade antibiotic treatment, or evade the host immune response. Four bovine-adapted lineages (CC71, CC97, CC151 and ST136) were identified among a collection of Irish S. aureus mastitis isolates. Genotypic variation of mastitis-causing strains may contribute to different presentations of the disease, including differences in milk somatic cell count (SCC), the main method of mastitis detection. The objective of this study was to investigate the influence of bacterial strain and lineage on host immune response, by employing cell culture methods in vitro as well as an in vivo infection model. Twelve bovine adapted S. aureus strains were examined for internalization into bovine mammary epithelial cells (bMEC) and their ability to induce an immune response from bMEC (using qPCR and ELISA). In vitro studies found differences in a variety of virulence traits between the lineages. Strains from lineages CC97 and CC71 internalized more efficiently into bovine mammary epithelial cells (bMEC) than CC151 and ST136. CC97 strains also induced immune genes in bMEC more strongly than strains from the other 3 lineages. One strain each of CC151 and CC97 that differed in their ability to cause an immune response in bMEC were selected on the basis of the above in vitro experiments. Fourteen first-lactation Holstein-Friesian cows were purchased from 2 farms on the basis of low SCC (less than 50 000 cells/ml) and infection free status. Seven cows were infected with 1.73 x 102 c.f.u. of the CC97 strain (Group 1) and another seven with 5.83 x 102 c.f.u. of the CC151 strain (Group 2). The contralateral quarter of each cow was inoculated with PBS (vehicle). Clinical signs of infection (temperature, milk and udder appearance, milk yield) were monitored for 30 days. Blood and milk samples were taken to determine bacterial counts in milk, SCC, white blood cell populations and cytokines. Differences in disease presentation in vivo between groups were observed, with two animals from Group 2 developing clinical mastitis and requiring antibiotic treatment, while one animal from Group 1 did not develop an infection for the duration of the study. Fever (temperature > 39.5⁰C) was observed in 3 animals from Group 2 and in none from Group 1. Significant differences in SCC and bacterial load between groups were observed in the initial stages of infection (week 1). Data is also being collected on cytokines and chemokines secreted during the course of infection. The results of this study suggest that a strain from lineage CC151 may cause more severe clinical mastitis, while a strain from lineage CC97 may cause mild, subclinical mastitis. Diversity between strains of S. aureus may therefore influence the clinical presentation of mastitis, which in turn may influence disease detection and treatment needs.

Keywords: Bovine mastitis, host immune response, host-pathogen interactions, Staphylococcus aureus

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Authors: Cherity A. Ysquierdo, Pia U. Olafson, Donald B. Thomas

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Musca domestica L. were collected from cattle diagnosed with bovine ringworm to evaluate the potential of the house fly to disseminate Trichophyton verrucosum E. Bodin, a fungal dermatophyte that is the causative agent for ringworm in cattle. Fungal isolates were cultured from 45 individual flies on supplemented Sabouraud dextrose agar, and isolates were identified using morphological and microscopic approaches. Each isolate was further identified by PCR amplification of the ribosomal DNA locus with fungal specific primers and subsequent amplicon sequencing. No T. verrucosum were identified using these approaches. However, 36 different fungal species representing 17 genera were cultured from these flies, including several allergenic and pathogenic species. Several species within the fungal orders Hypocreales, Microascales, Onygenales, Saccharomycetales, Xylaniales, and Agaricales were observed for the first time on house flies. The most frequent fungus recovered was Cladosporium cladosporoides, which is known to be a ubiquitous, airborne allergen.

Keywords: bovine ringworm, Cladosporium, dermatophyte, Musca domestica

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Authors: Mahfoodha Al-Kitani, Dylan Thompson, Keith Stokes

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Sickle cell disease (SCD) and sickle cell trait (SCT) are the most common hematological diseases in Oman according to the national survey of genetic blood disorders. The aim of this study was to determine markers of physical fitness and anthropometrics indices in children with sickle cell disease and children with sickle cell trait and compare them with normal healthy children of the same age. One hundred and twenty male children participated in the present study divided to three groups: 40 with sickle disease (SCD; age, 13.3(.80), height, 131.9(3.5), mass, 29.2(3.1)); 40 with sickle cell trait (SCT; age, 12.2(.80), height, 141.0(9.9), mass, 38.0(4.4)); and 40 controls with normal hemoglobin (Con; age, 12.8(.80), height, 139.4(8.7), mass, 37.2(4.3)). All children completed a 5-min running exercise test on a treadmill at speed corresponding to 5 km/hr. Heart rate and was recorded during exercise and during 10-min of recovery. Blood lactate was measured before and 5 min after the completion of exercise. Children with SCD exhibited a higher mean value (P < 0.05) for percent body fat and fat mass than the normal healthy subjects and SCT subjects. Resting values of hemoglobin were similar in SCT (11.04(.78)) and control (10.8(94)) groups, and lower in SCD (8.89(.54); P < 0.05). There was a strong correlation between peak heart rate and resting hemoglobin levels for the three groups (r= -.472. n= 120, p < .0005).The SCD group (175.2(10.3)) exhibited higher mean heart rate during exercise than those observed in the SCT (143.7(9.5)) and normal control children (144.5(22.4); P < 0.05). Additionally, SCD children showed higher serum lactate values before and after treadmill exercise compared to the other groups (P < 0.05). Children with sickle cell trait demonstrate similar physical fitness level and similar exercise responses to treadmill stress test to normal children. In contrast, SCD children have lower body mass, higher fat mass and lower physical fitness than children with SCT and healthy controls.

Keywords: sickle cell disease, sickle cell trait, children, exercise

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Authors: Ismail Bayram, Aamir Iqbal, E. Eren Gultepe, Cangir Uyarlar, Umit Ozcınar, I. Sadi Cetingul

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The current study was planned with the objective to explore the potential of pomegranate molasses (PM) on performance, egg quality and blood parameters in older laying hens. A total of 240 Babcock white laying hens (52 weeks old) were divided into 5 groups (n=48) with 8 subgroups having 6 hens in each. Pomegranate molasses was added in the drinking water to experimental groups with 0 %, 0.1%, 0.25 %, 0.5%, and 1%, respectively during one month. In our results, egg weight values were remained the same in all pomegranate molasses supplemented groups except 1% group over control. However, feed consumption, egg production, feed conversion ratio (FCR), egg mass, egg yolk cholesterol, body weights, and water consumption remained unaffected (P > 0.05). During mid-study (15 Days) analyses, egg quality parameters such as Haugh unit, eggshell thickness, albumin index, yolk index, and egg yolk color were remained non-significant (P > 0.05) while after final (30 Days) egg analyses, only egg yolk color had positively (P < 0.05) increased in 0.5% group. Moreover, Haugh unit, eggshell thickness, and albumin index were not significantly (P > 0.05) affected by the supplementation of pomegranate molasses. Regarding serological parameters, pomegranate molasses did not show any positive effect on cholesterol, total protein, LDL, HDL, GGT, AST, ALT, and glucose level. Similarly, pomegranate molasses also showed non-significant (P > 0.05) results on different blood parameters such as HCT, RBC, MCV, MCH, MCHC, PLT, RDWC, MPV except hemoglobin level. Only hemoglobin level was increased in all experimental groups over control showing that pomegranate molasses can be used as an enhancer in animals with low hemoglobin level.

Keywords: pomegranate molasses, laying hen, egg yield, blood parameters

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Authors: Amanda Cristina de Oliveira, Elias de Souza Monteiro Filho, Roberta Ceriani

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Liquid-liquid extraction in aqueous two-phase systems (ATPSs) constitutes a powerful tool for purifying bio-materials, such as cells, organelles, proteins, among others. In this work, the extraction of the bovine serum albumin (BSA) has been studied in systems formed by polyethylene glycol (PEG) (1500, 4000, and 6000 g.mol⁻¹) + potassium sodium tartrate + water at 303.15°K. Phase diagrams were obtained by turbidimetry and Merchuk’s method (1998). The experimental tie-lines were described using the Othmer-Tobias and Bancroft correlations. ATPSs were correlated with the nonrandom two-liquid (NRTL) model. The results were considered excellent according to global root-mean-square deviations found which were between 0,72 and 1,13%. The concentrations of the proteins in each phase were determined by spectrophotometry at 280 nm, finding partition efficiencies greater than 71%.

Keywords: aqueous two phases systems, bovine serum albumin , liquid-liquid extraction, polyethylene glycol

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Authors: Fasil Wagnew, Kefyalew Addis Alene, Setegn Eshetie, Tom Wingfield, Matthew Kelly, Darren Gray

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Introduction: Undernutrition is a major and under-appreciated risk factor for TB, which is estimated to be responsible for 1.9 million TB cases per year globally. The effectiveness of micronutrient supplementation on TB treatment outcomes and its prognostic markers such as sputum conversion and serum zinc, retinol, and hemoglobin levels has been poorly understood. This systematic review and meta-analysis aimed to determine the association between zinc and vitamin A supplementation and TB treatment outcomes and its prognostic markers. Methods: A systematic literature search for randomized controlled trials (RCTs) was performed in PubMed, Embase, and Scopus databases. Meta-analysis with a random effect model was performed to estimate risk ratio (RR) and mean difference (MD), with a 95% confidence interval (CI), for dichotomous and continuous outcomes, respectively. Results: Our search identified 2,195 records. Of these, nine RCTs consisting of 1,375 participants were included in the final analyses. Among adults with pulmonary TB, zinc (RR: 0.94, 95%CI: 0.86, 1.03), vitamin A (RR: 0.90, 95%CI: 0.80, 1.01), and combined zinc and vitamin A (RR: 0.98, 95%CI: 0.89, 1.08) supplementation were not significantly associated with TB treatment success. Combined zinc and vitamin A supplementation was significantly associated with increased sputum smear conversion at 2 months (RR: 1.16, 95%CI: 1.03, 1.32), serum zinc levels at 2 months (MD of 0.86umol/l, 95% CI: 0.14, 1.57), serum retinol levels at 2 months (MD: 0.06umol/l, 95 % CI: 0.04, 0.08) and 6 months (MD: 0.12umol/l, 95 % CI: 0.10, 0.14), and serum hemoglobin level at 6 months (MD: 0.29 ug/dl, 95% CI: 0.08 to 0.51), among adults with TB. Conclusions: Providing zinc and vitamin A supplementation to adults with pulmonary TB during treatment may increase early sputum smear conversion, serum zinc, retinol, and hemoglobin levels. However, the use of zinc, vitamin A, or both were not associated with TB treatment success.

Keywords: zinc and vitamin A supplementation, tuberculosis, treatment outcomes, meta-analysis, RCT

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Authors: Mannan Malik Muhammad Naeem, Jeong Myung Yung

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Functional near infrared spectroscopy (fNIRS) has a favorable position in non-invasive brain imaging techniques. The concentration change of oxygenated hemoglobin and de-oxygenated hemoglobin during particular cognitive activity is the basis for this neuro-imaging modality. Two wavelengths of near-infrared light can be used with modified Beer-Lambert law to explain the indirect status of neuronal activity inside brain. The temporal resolution of fNIRS is very good for real-time brain computer-interface applications. The portability, low cost and an acceptable temporal resolution of fNIRS put it on a better position in neuro-imaging modalities. In this study, an optimization model for impulse response function has been used to estimate/predict initial dip using fNIRS data. In addition, the activity strength parameter related to motor based cognitive task has been analyzed. We found an initial dip that remains around 200-300 millisecond and better localize neural activity.

Keywords: fNIRS, brain-computer interface, optimization algorithm, adaptive signal processing

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Authors: Athar Hussain Memon

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Objectives: We tried to determine the frequency of raised C-reactive protein (CRP) in patients with type 2 diabetes mellitus. Patients and Methods: This cross-sectional descriptive study of six months study was conducted at Liaquat University Hospital Hyderabad from March 2013 to August 2013. All diabetic patients of ≥35 years age of either gender for >01 year duration visited at OPD were evaluated for C-reactive protein and their glycemic status by hemoglobin A1c. The data was analyzed in SPSS and the frequency and percentage were calculated. Results: During six month study period, total 100 diabetic patients were evaluated for C-reactive protein. The majority of patients were from urban areas 75/100 (75%). The mean ±SD for age of patients with diabetes mellitus was 51.63±7.82. The mean age ±SD of patient with raised CRP was 53±7.21. The mean ±SD for HbA1c in patients with raised CRP is 9.55±1.73. The mean random blood sugar level in patients with raised CRP was 247.42 ± 6.62. The majority of subjects were of 50-69 years of age group with female predominance (p=0.01) while the CRP was raised in 70 (70%) patients in relation to age (p=0.02) and gender (p=0.01), respectively. Both HbA1c and CRP were raised in 64.9% (p=0.04) in patients with type 2 diabetes mellitus. The mean ±SD of CRP was 5.8±1.21 while for male and female individuals with raised CRP was 3.52±1.22 and 5.7±1.63, respectively. Conclusions: The raised CRP was observed in patients with type 2 diabetes mellitus.

Keywords: diabetes mellitus, C-reactive protein, hemoglobin A1c, diabetes and metabolism

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Authors: Shu Hwang Ang, Choo Yee Yu, Geik Yong Ang, Yean Yean Chan, Yatimah Binti Alias, And Sook Mei Khor

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With the high prevalence of Type 2 diabetes mellitus across the world, the morbidities and mortalities associated with Type 2 diabetes have significant impact on the production line for a nation. With routine scheduled clinical visits to manage Type 2 diabetes, diabetic patients with hectic lifestyles can have low clinical compliance. Hence, it often decreases the effectiveness of diabetic management personalized for each diabetic patient. Here, we report a useful developed point-of-care (POC) device that detect glycated hemoglobin (HbA1c, biomarker for long-term Type 2 diabetic management). In fact, the established POC devices certified to be used in clinical setting are not only expensive ($ 8 to $10 per test), they also require skillful practitioners to perform sampling and interpretation. As a paper-based biosensor, the developed HbA1c biosensor utilized lateral flow principle to offer an alternative for cost-effective (approximately $2 per test) and end-user friendly device for household testing. Requiring as little as 2 L of finger-picked blood, the test can be performed at the household with just simple dilution and washings. With visual interpretation of numbers of test lines shown on the developed biosensor, it can be interpreted as easy as a urine pregnancy test, aided with scale of intensity provided. In summary, the developed HbA1c immunosensor has been tested to have high selectivity towards HbA1c, and is stable with reasonably good performance in clinical testing. Therefore, our developed HbA1c immunosensor has high potential to be an effective diabetic management tool to increase patient compliance and thus contain the progression of the diabetes.

Keywords: blood, glycated hemoglobin (HbA1c), lateral flow, type 2 diabetes mellitus

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Authors: Satwik Majumder, Dongyun Jung, Jennifer Ronholm, Saji George

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Bovine mastitis is the most common infectious disease in dairy cattle, with major economic implications for the dairy industry worldwide. Continuous monitoring for the emergence of antimicrobial resistance (AMR) among bacterial isolates from dairy farms is vital not only for animal husbandry but also for public health. In this study, the prevalence of AMR in 113 Escherichia coli isolates from cases of bovine clinical mastitis in Canada was investigated. Kirby-Bauer disk diffusion test with 18 antibiotics and microdilution method with three heavy metals (copper, zinc, and silver) was performed to determine the antibiotic and heavy-metal susceptibility. Resistant strains were assessed for efflux and ß-lactamase activities besides assessing biofilm formation and hemolysis. Whole-genome sequences for each of the isolates were examined to detect the presence of genes corresponding to the observed AMR and virulence factors. Phenotypic analysis revealed that 32 isolates were resistant to one or more antibiotics, and 107 showed resistance against at least one heavy metal. Quinolones and silver were the most efficient against the tested isolates. Among the AMR isolates, AcrAB-TolC efflux activity and ß-lactamase enzyme activities were detected in 13 and 14 isolates, respectively. All isolates produced biofilm but with different capacities, and 33 isolates showed α-hemolysin activity. A positive correlation (Pearson r = +0.89) between efflux pump activity and quantity of biofilm was observed. Genes associated with aggregation, adhesion, cyclic di-GMP, quorum sensing were detected in the AMR isolates, corroborating phenotype observations. This investigation showed the prevalence of AMR in E. coli isolates from bovine clinical mastitis. The results also suggest the inadequacy of antimicrobials with a single mode of action to curtail AMR bacteria with multiple mechanisms of resistance and virulence factors. Therefore, it calls for combinatorial therapy for the effective management of AMR infections in dairy farms and combats its potential transmission to the food supply chain through milk and dairy products.

Keywords: antimicrobial resistance, E. coli, bovine mastitis, antibiotics, heavy-metals, efflux pump, ß-lactamase enzyme, biofilm, whole-genome sequencing

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Authors: Sanjeev Kumar, Swati Choudhary

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Background: A variety of studies warn that sedentary lifestyles can contribute to many preventable causes of death. This study was taken to determine the effects of two types of aerobic training programs on erythrocytes, leukocytes, hemoglobin concentration (Hb), platelets and hematocrit of sedentary people (N=60) with age group 20 to 30 years. Methods: All the subjects were randomly divided into three groups i.e. two experiments groups (aerobic dance & cardio fitness) and control group. Each group having 10 male and 10 females. Experimental groups undergone 60 minutes of training 5 times a week for 12 weeks whereas the control group did not participate in any training program except their daily routine. The aerobic dance group was chosen to perform exercise like step –touch, side-to-side, V-step and hand and body movements, etc. The cardio fitness group was chosen to perform exercises with modern fitness equipment like treadmill, elliptical trainer, stationary bike and rowing machine. Rating of perceived exertion (RPE) scale developed by Gunner Borg was used to monitor the intensity of the workout. Aerobic programs were encompassed of low-impact (0- 4 week & perceived exertion from 6 to 12), moderate-impact (4-8 week and perceived exertion from 12 to 16) and high-impact (8- 12 week & perceived exertion from 16 to 20). Results: To test the effectiveness of training programs paired t-test was used and significant difference (p<0.05) was observed in erythrocytes, hemoglobin concentration, platelets, hematocrit but no significant effects of training was found in leukocytes (p>0.05). Paired t-test also showed that no effect of time was seen in the control group in all the cases (p>0.05). Further analysis of covariance was used to know which program was more effective and it was seen that F value was found significant in the case of erythrocytes, hemoglobin concentration, platelets, and hematocrit as their associated p-value (p<0.05) is lesser than 0.05. As F value was found significant for hematological parameters, fishers least significant difference test was used and results of post hoc mean comparison indicated that experimental groups (aerobic dance group and cardio fitness group) had significant difference with control group in erythrocytes, hemoglobin concentration, platelets and hematocrit and insignificant difference was found between aerobic dance group & cardio fitness group in all the cases. Thus, it may be concluded that in general, both the aerobic training programs had adequate effects on all the hematological parameters except leukocytes.

Keywords: aerobic dance, cardio fitness, hematological variables, rating perceived exertion scale

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Authors: Sibel Daglilar, Isil Kerti, Murat Karagoz, Fatih Dumludag, Oguzhan Gunduz, Faik Nuzhet Oktar

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Hydroxyapatite (HA) materials have common use in bone repairing due to its ability to accelerate the bone growth around the implant. In spite of being a biocompatible and bioactive material, HA has a limited usage as an implant material because of its weak mechanical properties. HA based composites are required to improve the strength and toughness properties of the implant materials without compromising of biocompatibility. The excellent mechanical properties and higher biocompatibilities are expected from each of biomedical composites. In this study, HA composites were synthesized by using bovine bone reinforced doped with different amount of (wt.%) Li2O. The pressed pellets were sintered at various sintering temperatures between 1000ºC and 1300°C, and mechanical, electrical properties of the obtained products were characterized. In addition to that, in vitro stimulated body fluid (SBF) tests for these samples were conducted. The most suitable composite composition for biomedical applications was discussed among the composites studied.

Keywords: biocomposites, sintering temperature, biocompatibility, electrical property, conductivity, mechanical property

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Authors: Sri Wahjuningsih, Nur Ihsan, Hadiah

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In the embryo transfer program, to address the limited production of embryos in vivo, in vitro embryo production has become an alternative approach that is relatively inexpensive. One potential source of embryos that can be developed is to use immature oocytes then conducted in vitro maturation and in vitro fertilization. However, obstacles encountered were oocyte viability mammals have very limited that it cannot be stored for a long time, so we need oocyte cryopreservation. The research was conducted to know the optimal concentration use of ethylene glycol as a cryoprotectant on oocytes freezing.Material use in this research was immature oocytes; taken from abbatoir which was aspirated from follicle with diameter 2-6 mm. Concentration ethylen glycol used were 0,5 M, I M, 1,5 M and 2M. The freezing method used was conventional method combined with a five-step protocol washing oocytes from cryoprotectant after thawing. The result showed that concentration ethylen glycol have the significant effect (P<0.05) on oocytes quality after thawing and in vitro maturation. It was concluded that concentration 1,5 M was the best concentration for freezing oocytes using conventional method.

Keywords: bovine, conventional freezing, ethylen glycol, oocytes

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Authors: Cigdem Gul, Murat Tosunoglu, Nurcihan Hacioglu

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The contamination or desiccation of fresh waters also has a negative effect on freshwater turtles like other fresh water-dependent species. In order to conserve those species, which are confronted with such negative conditions, it is necessary to know beforehand the biology and the physiology of species. In this study, a comprehensive health assessment was conducted on a total of 20 clinically normal individuals free living Western Caspian Turtle (Mauremys rivulata) captured from two different freshwater localities in the Biga stream (Çanakkale-Turkey). When comparing our findings with the Turkish legislation (Water pollution control regulation), the 1. Locality of the Biga stream in terms of total coliform classified as "high quality water" (Coliform: 866.66 MPN/100 mL), while the 2. Locality was a “contaminated water" (Coliform: 53266.66 MPN/100 mL). Blood samples for hematological and biochemical analyses were obtained from the dorsal coccygeal vein. A total of 1-2 mL of blood was collected from each of the specimens via needle. After the required procedures had been performed, the turtles were put back in the same localities. Hematological and biochemical analyses based on high quality water and contaminated water, respectively, are as follows: Red blood cell count (512600-582666.66 per cubic millimeter of blood), white blood cell count (5920-5980 per cubic millimeter of blood), hematocrit value (24-24.66 %), hemoglobin concentration (6.52-6.35 g/dl), mean corpuscular volume (466.20-468.98 fl), mean corpuscular hemoglobin (125.77-113.84 pg), mean corpuscular hemoglobin concentration (28.25-26.49 %), glucose (94.43-87.43 mg/dl), creatinine (0.23-0.3241 mg/dl), uric acid (12.59-10.48 mg/L), albumin (1.46-1.25 g/dl), calcium (8.67-9.59 mg/dl), triglyceride (95.55-75.21 mg/dl), and total protein (4.85-3.45 g/dl). When an examination was made depending on the water quality of freshwater, variations were detected in hematology and biochemistry values, but not found significant difference.

Keywords: biochemistry, freshwater quality, hematological parameters, Mauremys rivulata.

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Authors: Forough Ataollahi, Sumit Pramanik, Belinda Pingguan-Murphy, Wan Abu Bakar Bin Wan Abas, Noor Azuan Bin Abu Osman

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Endothelium proliferation is an important process in cardiovascular homeostasis and can be regulated by extracellular environment, as cells can actively sense mechanical environment. In this study, we evaluated endothelial cell proliferation on PDMS/alumina (Al2O3) composites and pure PDMS. The substrates were prepared from pure PDMS and its composites with 5% and 10% Al2O3 at curing temperature 50˚C for 4 h and then characterized by mechanical, structural and morphological analyses. Higher stiffness was found in the composites compared to the pure PDMS substrate. Cell proliferation of the cultured bovine aortic endothelial cells on substrate materials were evaluated via Resazurin assay and 1, 1’-Dioctadecyl-1, 3, 3, 3’, 3’-Tetramethylindocarbocyanine Perchlorate-Acetylated LDL (Dil-Ac-LDL) cell staining, respectively. The results revealed that stiffer substrates promote more endothelial cells proliferation to the less stiff substrates. Therefore, this study firmly hypothesizes that the stiffness elevates endothelial cells proliferation.

Keywords: stiffness, proliferation, bovine aortic endothelial cells, extra cellular matrix, vascular

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Authors: Alhaji Musa Abdullahi, Usman Abdullahi, Adamu Lawan, Aminu Maidala

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Abstract 16 healthy lactating cows will be randomly selected for the trial and will be randomly divided in to 4 groups with 4 cows in each. They will be kept under similar management condition (conventional management system). Animals of relatively same weight and age will be used. After 11days for adaptation, feed intake and performance of the experimental animals will be determine. Milk sample will be collected at each milking in the morning and afternoon to determine; Milk yield, Milk fat percentage, Solid not fat percentage, Total solid percentage of milk. Cows dung will be observe to determine; Score 1 very loose watery stool, Score 2 semi solid with undigested raw material, Score 3 semi solid with less undigested raw material, Score 4 solid with very less undigested raw material, Score 5 good dung no undigested raw material. At the end of the experiment, blood samples will be analyzed for full blood counts and differentials {White Blood Cells (WBC), Red Blood Cells (RBC), Hemoglobin (Hb), Packed Cell Volume (PCV), Mean Corpuscular Volume (MCV), Mean Corpuscular Hemoglobin (MCH), Mean Corpuscular Hemoglobin Concentration (MCHC), Platelets (PLT), Lymphocytes (LYM), Basophils, Eosinophils and Monocytes Proportion (MXD) and Neutrophils (NEUT)} using automated hematology analyzer. Serum samples will be analyzed for heat shock transcription factors, heat shock proteins and hormones (Serum glucocorticoid, prolactin and cortisol). Moreover, biochemical analysis will also be conducted to check for Total protein (TP), Albumen (ALB), Globulin (GBL), Total cholesterol (TCH), glucose (G), sodium (Na+), potassium (K+), chloride (Cl-) and pH. Keywords: Lactating cows, milk composition, dung score and blood parameters.

Keywords: Lactating cows , Milk yield , Dung score , Blood parameters

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Authors: Fatima Abu Baker Hamad

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Anaemia is a public health problem that affects population in both rich and poor countries. Although the primary cause is iron deficiency, it is seldom present in isolation. More frequently it coexists with a number of other causes, such as malaria, parasitic infection, nutritional deficiencies and hemoglobin apathies. That was the people in Sudan suffered from it .Anaemia has a high prevalence in patients with cancer. The aim of this study was to find the incidence of anaemia in new cases of Sudanese female breast patients attending the National Cancer Institute (NCI), Gezira University, Sudan. The study was performed on 250 female breast cancer patients, the age range was (20-70) years and the mean age was 45.99±0.82. The hemoglobin level was measured by SYSMEX-KX2lM.As result 144(58.8) of patients presented with anaemia, between moderate to severe. Forty four (17.6%) of the patients were found to be under weight, 31 of them were anaemic. While 105(42%) of the patients were overweight and obese, 52 of them were anaemic. The incidence of anaemia in newly diagnosed Sudanese female breast cancer patients presented at NCI is association presentation with advance disease stage. Also it is related to age, state of nutrition and social economic factors. Early cancer detection which leads to effective treatment and reduced complication of diseases included anaemia is recommended.

Keywords: anaemia, breast cancer, stages of disease, malaria

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Authors: Siyanbola Mojisola Funmilayo, Amao Emmanuel Ayodele

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The poultry industry in Nigeria has been played by a variety of problems, which include the search for feed ingredients that are not competed for by man. This has resulted in a reduced interest of farmers in the industry leading to a reduction in animal protein availability for human consumption as a consequence of a high cost of production. The incorporation of wild sunflower meal (Tithonia diversfolia, Hemsl A. Gray) (WSF Meal) and some others in poultry diets have been reported to result in compounded feed with nutrient profiles that compare favourable with feeds of conventional feedstuff and reduce feed cost as they reduce competition with humans. A 98-day feeding trial was used to evaluate the effect of Wild sunflower leaf (WSL) at varying levels on the hematology and biochemistry of cockerels. A total of one hundred and twenty(120) cockerel birds were randomly allotted into four experimental diets with three replicates per experimental diet (ten birds per replicate). Wild sunflower leaf was included in four graded levels ; 0, 5, 10, and 15%. Packed cell volume, Red blood cell count, White blood cell count, Hemoglobin count, Lymphocyte count, Neutrophil count, Platelets, Mean Corpuscular Hemoglobin Concentration (MCHC), Mean Corpuscular Hemoglobin (MCH), Aspartate aminotransferase (AST), Glucose, Urea, Chloride, Sodium, and Potassium ion values were significantly different (p<0.05) among the treatments. Mean values obtained for Creatinine, Total Protein, Alanine aminotransferase (ALT), Albumin, and Mean Corpuscular Volume (MCV) were not significantly different (p>0.05) in all the treatment. WSL could be included up to 15% in the diet of cockerel without any deleterious effect on the birds. Based on the results, up to 15% Wild sunflower meal (WSL) can be included in the diet of cockerel without any adverse effect on the hematology and biochemical indices of birds.

Keywords: biochemical changes, cockerels, hematology, wild sunflower leaf

Procedia PDF Downloads 447

Authors: Mukesh N. Kher, Anju P. Kunjadia, Dev S. Nauriyal, Chaitanya G. Joshi, Navin R. Sheth, Vaibhav D. Bhatt

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In-vivo immunotherapeutic potential on cytokines production and antibacterial activity of a topical herbal gel was evaluated in two breeds of cattle in bovine subclinical mastitis. The response to treatment was evaluated by enumerating somatic cell count (SCC), determining total bacterial count and studying the expression of different cytokines like (interleukin 6, 8, 12, GMCSF, interferon–γ and TNF‑α). The pre‑ and post‑treatment SCC in mastitic quarters did not differ statistically-significantly. However, total bacterial count declined significantly from day 0 onwards in both the breeds. Significant differences (P < 0.01) were observed in all types of cytokines production on day 0, 5, and 21 post last treatments in both the breeds. The comparison of cytokine expression profiles between crossbred and Gir cattle affirmed a significant difference in expression of IL-6 and TNF-α. The topical herbal gel showed immunomodulatory and antimicrobial activities in subclinical mastitis, and therefore the work supports its use as substitute herbal therapy against subclinical mastitis in bovines.

Keywords: antibacterial activity, immunomodulation, herbal gel, subclinical mastitis

Procedia PDF Downloads 288

Authors: Eva Tvrdá, Boris Botman, Marek Halenár, Tomáš Slanina, Norbert Lukáč

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In vitro storage and processing of animal semen represents a risk factor to spermatozoa vitality, potentially leading to reduced fertility. A variety of substances isolated from natural sources may exhibit protective or antioxidant properties on the spermatozoon, thus extending the lifespan of stored ejaculates. This study compared the ability of different concentrations of the Salvia officinalis extract on the motility, mitochondrial activity, viability and reactive oxygen species (ROS) production by bovine spermatozoa during different time periods (0, 2, 6 and 24 h) of in vitro culture. Spermatozoa motility was assessed using the Computer-assisted sperm analysis (CASA) system. Cell viability was examined using the metabolic activity MTT assay, the eosin-nigrosin staining technique was used to evaluate the sperm viability and ROS generation was quantified using luminometry. The CASA analysis revealed that the motility in the experimental groups supplemented with 0.5-2 µg/mL Salvia extract was significantly lower in comparison with the control (P<0.05; Time 24 h). At the same time, a long-term exposure of spermatozoa to concentrations ranging between 0.05 µg/mL and 2 µg/mL had a negative impact on the mitochondrial metabolism (P<0.05; Time 24 h). The viability staining revealed that 0.001-1 µg/mL Salvia extract had no effects on bovine male gametes, however 2 µg/mL Salvia had a persisting negative effect on spermatozoa (P<0.05). Furthermore 0.05-2 µg/mL Salvia exhibited an immediate ROS-promoting effect on the sperm culture (P>0.05; Time 0 h and 2 h), which remained significant throughout the entire in vitro culture (P<0.05; Time 24 h). Our results point out to the necessity to examine specific effects the biomolecules present in Salvia officinalis may have individually or collectively on the in vitro sperm vitality and oxidative profile.

Keywords: bulls, CASA, MTT test, reactive oxygen species, sage, Salvia officinalis, spermatozoa

Procedia PDF Downloads 338

Authors: Van Huong Do, Simon Walton, German Amaya, Madeline Batsiokis, Sally Catt, Andrew Taylor-Robinson

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Identification of the most suitable stages of bovine in vitro-derived blastocysts (early, expanded and hatching) prior to vitrification is a straightforward process that facilitates the decision as to which blastocyst stage to use for transfer of fresh and vitrified embryos. Research on in vitro evaluation of suitable stages has shown that the more advanced developmental stage of blastocysts is recommended for fresh embryo transfer while the earlier stage is proposed for embryo transfer following vitrification. There is, however, limited information on blastocyst stages using in vivo assessment. Hence, the aim of the present study was to determine the optimal stage of a blastocyst for vitrification and embryo transfer through a two-step procedure of embryo transfer followed by pregnancy testing at 35, 60 and 90 days of pregnancy. 410 good quality oocytes aspirated by the ovum pick-up technique from 8 donor cows were subjected to in vitro embryo production, vitrification and embryo transfer. Good quality embryos were selected, subjected to vitrification and embryo transfer. Subsequently, 77 vitrified embryos at different blastocyst stages were transferred to synchronised recipient cows. The overall cleavage and blastocyst rates of oocytes were 68.8% and 41.7%, respectively. In addition, the fertility and blastocyst production of 6 bulls used for in vitro fertilization was examined and shown to be statistically different (P<0.05). Results of ongoing pregnancy trials conducted at 35 days, 60 days and 90 days will be discussed. However, preliminary data indicate that individual bulls demonstrate distinctly different fertility performance in vitro. Findings from conception rates would provide a useful tool to aid selection of bovine in vitro-derived embryos for vitrification and embryo transfer in commercial settings.

Keywords: blastocyst, embryo transfer, in vitro-derived embryos, ovum pick-up, vitrification

Procedia PDF Downloads 308

Authors: A. Bhattacharya, Sanjay Gupta, S. H. Bodakhe

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HbA1c is form of the haemoglobin that is used to measure the average plasma glucose concentration over prolonged periods of time. It is formed in a non-enzymatic glycation pathway by hemoglobin's exposure to plasma glucose. In diabetes mellitus, higher amounts of glycated hemoglobin, indicating poorer control of blood glucose levels, have been associated with cardiovascular disease, nephropathy, and retinopathy. Guraku’s basic components are nicotine and jaggery, jaggery is made up of sugarcane so can have a diabetogenic potential which is exacerbated in presence of nicotine. This work had done with the aim to find correlation between Diabetic cataract, HbA1c and Guraku. Subjects were enrolled according to the inclusion and exclusion criteria. In this study total 75 subjects were included. In the study it was found that people consuming Guraku had a high level of HbA1c thus are more prone to the development of diabetic cataract. Male subjects are the more than female subjects. Most of the subjects belong to the lower socioeconomical class and not very educated. It could be concluded that this type of study could be useful in indentifying number of subjects suffering from diabetic cataract whose condition get worse by use of nicotine product like Guraku and preventive measure to be taken in prevention of this type of diabetic complication.

Keywords: diabetic cataract, HbA1c, Guraku, diabetogenic potential

Procedia PDF Downloads 403

Authors: Claudivan Costa de Lima, Angelo da Silva Monteiro

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With the aim of evaluating the effects of grounding with copper wire on the reduction of the microbial load of biocontaminated milk samples and on their acidification over time, two complementary experiments were carried out. In the first, the treatments consisted of: i) raw milk sample (control), ii) slow pasteurization, iii) grounding with copper wire and, iv) contact with copper ring. Analyzes of total, thermoresistant and mesophilic coliforms were performed 30 minutes after the application of these treatments. In the second experiment, under the same conditions as the first, measurements of pH and Dornic acidity were performed at 0, 0.5, 2, 4, 8, 12, and 24 h from the installation of the experiment. Pasteurization eliminated almost all groups of bacteria present in the milk samples while grounding only allowed reductions in the population of thermotolerant coliforms and mesophiles, both greater than 95%, maintaining, however, unchanged the amounts of total coliforms. The copper ring, in turn, had no effect on the microbiological parameters studied. The reduction in the population of mesophiles in grounded milk samples, contrary to what happened with pasteurized milk, was not enough to inhibit the acidification process over the experimental period.

Keywords: pasteurization, low frequency electric current, thermotolerant coliforms, mesophiles in bovine milk

Procedia PDF Downloads 109

Authors: Olufemi M. Alabi, Foluke A. Aderemi, Adebayo A. Adewumi, Banwo O. Alabi

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General well-being of animals is of paramount interest in some developed countries and of global importance hence the shift onto alternative housing systems for egg-type chickens as replacement for conventional battery cage system. However, there is paucity of information on the effect of this shift on physiological status of the hens to judge their health via the blood profile. Therefore, investigation was carried out on two strains of hen kept in three different housing systems in humid tropics to evaluate changes in their blood parameters. 108, 17-weeks old super black (SBL) hens and 108, 17-weeks old super brown (SBR) hens were randomly allotted to three different intensive systems Partitioned Conventional Cage (PCC), Extended Conventional Cage (ECC) and Deep Litter System (DLS) in a randomized complete block design with 36 hens per housing system, each with three replicates. The experiment lasted 37 weeks during which blood samples were collected at 18th week of age and bi-weekly thereafter for analyses. Parameters measured are packed cell volume (PCV), hemoglobin concentration (Hb), red blood counts (RBC), white blood counts (WBC) and serum metabolites such as total protein (TP), albumin (Alb), globulin (Glb), glucose, cholesterol, urea, bilirubin, serum cortisol while blood indices such as mean corpuscular hemoglobin (MCH), mean cell volume (MCV) and mean corpuscular hemoglobin concentration (MCHC) were calculated. The hematological values of the hens were not significantly (p>0.05) affected by the housing system and strain, so also the serum metabolites except for the serum cortisol which was significantly (p<0.05) affected by the housing system only. Hens housed on PCC had higher values (20.05 ng/ml for SBL and 20.55 ng/ml for SBR) followed by hens on ECC (18.15ng/ml for SBL and 18.38ng/ml for SBL) while hens on DLS had the lowest value (16.50ng/ml for SBL and 16.00ng/ml for SBR) thereby confirming indication of stress with conventionally caged birds. Alternative housing systems can also be adopted for egg-type chickens in the humid tropics from welfare point of view with the results of this work confirming stress among caged hens.

Keywords: blood, housing, humid-tropics, layers

Procedia PDF Downloads 470

Authors: Befekadu Urga Wakayo

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Introduction: Bacterial infections represent major human and animal health problems in Ethiopia. In the face of poor antibiotic regulatory mechanisms, development of antimicrobial resistance (AMR) to commonly used drugs has become a growing health and livelihood threat in the country. Monitoring and control of AMR demand close coloration between human and veterinary services as well as other relevant stakeholders. However, risk of AMR transfer from animal to human population’s remains poorly explored in Ethiopia. This systematic research literature review attempted to give an overview on AMR challenges of bovine mastitis bacteria in Ethiopia. Methodology: A web based research literature search and analysis strategy was used. Databases are considered including; PubMed, Google Scholar, Ethiopian Veterinary Association (EVA) and Ethiopian Society of Animal Production (ESAP). The key search terms and phrases were; Ethiopia, dairy, cattle, mastitis, bacteria isolation, antibiotic sensitivity and antimicrobial resistance. Ultimately, 15 research reports were used for the current analysis. Data extraction was performed using a structured Microsoft Excel format. Frequency AMR prevalence (%) was registered directly or calculated from reported values. Statistical analysis was performed on SPSS – 16. Variables were summarized by giving frequencies (n or %), Mean ± SE and demonstrative box plots. One way ANOVA and independent t test were used to evaluate variations in AMR prevalence estimates (Ln transformed). Statistical significance was determined at p < 0.050). Results: AMR in bovine mastitis bacteria was investigated in a total of 592 in vitro antibiotic sensitivity trials involving 12 different mastitis bacteria (including 1126 Gram positive and 77 Gram negative isolates) and 14 antibiotics. Bovine mastitis bacteria exhibited AMR to most of the antibiotics tested. Gentamycin had the lowest average AMR in both Gram positive (2%) and negative (1.8%) bacteria. Gram negative mastitis bacteria showed higher mean in vitro resistance levels to; Erythromycin (72.6%), Tetracycline (56.65%), Amoxicillin (49.6%), Ampicillin (47.6%), Clindamycin (47.2%) and Penicillin (40.6%). Among Gram positive mastitis bacteria higher mean in vitro resistance was observed in; Ampicillin (32.8%), Amoxicillin (32.6%), Penicillin (24.9%), Streptomycin (20.2%), Penicillinase Resistant Penicillin’s (15.4%) and Tetracycline (14.9%). More specifically, S. aurues exhibited high mean AMR against Penicillin (76.3%) and Ampicillin (70.3%) followed by Amoxicillin (45%), Streptomycin (40.6%), Tetracycline (24.5%) and Clindamycin (23.5%). E. coli showed high mean AMR to Erythromycin (78.7%), Tetracycline (51.5%), Ampicillin (49.25%), Amoxicillin (43.3%), Clindamycin (38.4%) and Penicillin (33.8%). Streptococcus spp. demonstrated higher (p =0.005) mean AMR against Kanamycin (> 20%) and full sensitivity (100%) to Clindamycin. Overall, mean Tetracycline (p = 0.013), Gentamycin (p = 0.001), Polymixin (p = 0.034), Erythromycin (p = 0.011) and Ampicillin (p = 0.009) resistance increased from the 2010’s than the 2000’s. Conclusion; the review indicated a rising AMR challenge among bovine mastitis bacteria in Ethiopia. Corresponding, public health implications demand a deeper, integrated investigation.

Keywords: antimicrobial resistance, dairy cattle, Ethiopia, Mastitis bacteria

Procedia PDF Downloads 247

Authors: Sivakumar Singaravelu, Giriprasath Ramanathan, M. D. Raja, Uma Tirichurapalli Sivagnanam

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The goal of the present study was to develop and evaluate composite film for tissue engineering application. The keratin was extracted from bovine horn and used for preparation of keratin (HK), physiologically clotted fibrin (PCF) and gelatin (G) blend films in different stoichiometric ratios (1:1:1, 1:1:2 and 1:1:3) by using solvent casting method. The composite films (HK-PCF-G) were characterized physiochemically using Fourier Transform Infrared Spectroscopy (FTIR), Differential Scanning Calorimetry (DSC), Thermogravimetric Analysis (TGA) and Scanning Electron Microscopy (SEM). The mechanical properties of the composite films were analyzed. The results of tensile strength show that ultimate strength and elongation were 10.72 Mpa and 4.83 MPA respectively for 1:1:3 ratio combination. The SEM image showed a slight smooth surface for 1:1:3 ratio combination compared to other films. In order to impart antibacterial activities, the composite films were loaded with Mupirocin (MP) to act against infection. The composite films acted as a suitable carrier to protect and release the drug in a controlled manner. This developed composite film would be a suitable alternative material for tissue engineering application.

Keywords: bovine horn, keratin, fibrin, gelatin, tensile strength

Procedia PDF Downloads 450

Authors: Nabil Abu-Heakal, Ismail Abo-Ghanema, Basma Hamed Merghani

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The aim of this investigation is to study the effect of seasonal variations in Egypt on hematological parameters, reproductive and metabolic hormones of Egyptian buffalo-cows. This study lasted one year extending from December 2009 to November 2010 and was conducted on sixty buffalo-cows. Group of 5 buffalo-cows at estrus phase were selected monthly. Then, after blood sampling through tail vein puncture in the 2nd day after natural service, they were divided in two samples: one with anticoagulant for hematological analysis and the other without anticoagulant for serum separation. Results of this investigation revealed that the highest atmospheric temperature was in hot summer 32.61±1.12°C versus 26.18±1.67°C in spring and 19.92±0.70°C in winter season, while the highest relative humidity % was in winter season 43.50±1.60% versus 32.50±2.29% in summer season. The rise in temperature-humidity index from 63.73±1.29 in winter to 78.53±1.58 in summer indicates severe heat stress which is associated with significant reduction in total red blood cell count (3.20±0.15×106), hemoglobin concentration (8.83±0.43 g/dl), packed cell volume (30.73±0.12%), lymphocytes % (40.66±2.33 %), serum progesterone hormone concentration (0.56±0.03 ng/mll), estradiol17-B concentration (16.8±0.64 ng/ml), triiodothyronin (T3) concentration (2.33±0.33 ng/ml) and thyroxin hormone (T4) concentration (21.66±1.66 ng/ml), while hot summer resulted in significant increase in mean cell volume (96.55±2.25 fl), mean cell hemoglobin (30.81±1.33 pg), total white blood cell count (10.63±0.97×103), neutrophils % (49.66±2.33%), serum prolactin hormone (PRL) concentration (23.45±1.72 ng/ml) and cortisol hormone concentration (4.47±0.33 ng/ml) compared to winter season. There was no significant seasonal variation in mean cell hemoglobin concentration (MCHC). It was concluded that in Egypt there was a seasonal variation in atmospheric temperature, relative humidity, temperature humidity index (THI) and the rise in THI above the upper critical level (72 units), which, for lactating buffalo-cows in Egypt is the major constraint on buffalo-cows' hematological parameters and hormonal secretion that affects animal reproduction. Hence, we should improve climatic conditions inside the dairy farm to eliminate or reduce summer infertility.

Keywords: buffalo, climate change, Egypt, physiological parameters

Procedia PDF Downloads 662

Authors: M. H. Moreira da Silva, L. Valadao, F. Moreira da Silva

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In the present study, the fertilizing capacity of bovine spermatozoa was evaluated before and after its cryopreservation. For this, the testicles of 100 bulls slaughtered on Terceira Island were dissected, the epididymal tails were separated, and semen was recovered by the flotation method and then evaluated by phase contrast microscopy and by flow cytometry. For phase contrast microscopy, a drop of semen was used to evaluate the percentage of motile spermatozoa (from 0 to 100%) and motility (from 0 to 5). After determining the concentration and the abnormal forms, semen was diluted to a final concentration of 50 x 106 spz/ml and evaluated by flow cytometer for membrane and acrosome integrity using the conjugation of fluorescent probes propidium iodide (PI) and Arachis hypogea agglutinin (FITC-PNA). Freezing was carried out in a programmable semen freezer, using 0.25 ml straws, in a total of 20 x 106 viable sperm per straw with glycerol as a cryoprotectant in a final concentration of 0.58 M. It was observed that, on average, a total of 7.25 ml of semen was collected from each bull. The viability and vitality rates were respectively 83.22 ± 7.52% and 3.8 ± 0.4 before freezing, decreasing to 58.81 ± 11.99% and 3.6 ± 0.6, respectively, after thawing. Regarding cytoplasmic droplets, it was observed that a high percentage of spermatozoa had medial cytoplasmic droplets (38.47%), with only 3.32% and 0.15% presenting proximal and distal cytoplasmic drops, respectively. By flow cytometry, it was observed that before freezing, the percentage of sperm with the damaged plasma membrane and intact acrosome was 3.61 ± 0.99%, increasing slightly to 4.21 ± 1.86% after cryopreservation (p<0.05). Regarding spermatozoa with damaged plasma membrane and acrosome, the percentage before freezing was 3.37±1.87%, increasing to 4.34 ±1.16% after thawing, and no significant differences were observed between these two values. For the percentage of sperm with the intact plasma membrane and damaged acrosome, this value was 2.04 ± 2.34% before freezing, decreasing to 0.89 ± 0.48% after thawing (p<0.05). The percentage of sperm with the intact plasma membrane and acrosome before freezing was 90.99±2.75%, with a slight decrease to 90.57±3.15% after thawing (p<0.05). From this study, it can be clearly concluded that, after the slaughtering of bulls, the spermatozoa can be recovered from the epididymis and cryopreserved, maintaining an excellent rate of sperm viability and quality after thawing.

Keywords: bovine semen, epididymis, cryopreservation, fertility assessment

Procedia PDF Downloads 91