Search results for: frozen bivalve molluscs

Authors: Eleonora Di Salvo, Antonio Panebianco, Graziella Ziino

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Background: Vibrio parahaemolyticus is a human pathogen that is widely distributed in marine environments. It is frequently isolated from raw seafood, particularly shellfish. Consumption of raw or undercooked seafood contaminated with V. parahaemolyticus may lead to acute gastroenteritis. Vibrio spp. has excellent resistance to low temperatures so it can be found in frozen products for a long time. Recently, the viable but non-culturable state (VBNC) of bacteria has attracted great attention, and more than 85 species of bacteria have been demonstrated to be capable of entering this state. VBNC cells cannot grow in conventional culture medium but are viable and maintain metabolic activity, which may constitute an unrecognized source of food contamination and infection. Also V. parahaemolyticus could exist in VBNC state under nutrient starvation or low-temperature conditions. Aim: The aim of the present study was to optimize methods and investigate V. parahaemolyticus VBNC cells and their presence in frozen bivalve molluscs, regularly marketed. Materials and Methods: propidium monoazide (PMA) was integrated with real-time polymerase chain reaction (qPCR) targeting the tl gene to detect and quantify V. parahaemolyticus in the VBNC state. PMA-qPCR resulted highly specific to V. parahaemolyticus with a limit of detection (LOD) of 10-1 log CFU/mL in pure bacterial culture. A standard curve for V. parahaemolyticus cell concentrations was established with the correlation coefficient of 0.9999 at the linear range of 1.0 to 8.0 log CFU/mL. A total of 77 samples of frozen bivalve molluscs (35 mussels; 42 clams) were subsequently subjected to the qualitative (on alkaline phosphate buffer solution) and quantitative research of V. parahaemolyticus on thiosulfate-citrate-bile salts-sucrose (TCBS) agar (DIFCO) NaCl 2.5%, and incubation at 30°C for 24-48 hours. Real-time PCR was conducted on homogenate samples, in duplicate, with and without propidium monoazide (PMA) dye, and exposed for 45 min under halogen lights (650 W). Total DNA was extracted from cell suspension in homogenate samples according to bolliture protocol. The Real-time PCR was conducted with species-specific primers for V. parahaemolitycus. The RT-PCR was performed in a final volume of 20 µL, containing 10 µL of SYBR Green Mixture (Applied Biosystems), 2 µL of template DNA, 2 µL of each primer (final concentration 0.6 mM), and H2O 4 µL. The qPCR was carried out on CFX96 TouchTM (Bio-Rad, USA). Results: All samples were negative both to the quantitative and qualitative detection of V. parahaemolyticus by the classical culturing technique. The PMA-qPCR let us individuating VBNC V. parahaemolyticus in the 20,78% of the samples evaluated with a value between the Log 10-1 and Log 10-3 CFU/g. Only clams samples were positive for PMA-qPCR detection. Conclusion: The present research is the first evaluating PMA-qPCR assay for detection of VBNC V. parahaemolyticus in bivalve molluscs samples, and the used method was applicable to the rapid control of marketed bivalve molluscs. We strongly recommend to use of PMA-qPCR in order to identify VBNC forms, undetectable by the classic microbiological methods. A precise knowledge of the V.parahaemolyticus in a VBNC form is fundamental for the correct risk assessment not only in bivalve molluscs but also in other seafood.

Keywords: food safety, frozen bivalve molluscs, PMA dye, Real-time PCR, VBNC state, Vibrio parahaemolyticus

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Authors: Orawan Vichiansan, J. Kraipoj, K.Phandech, P. Sirasaporn

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Generally, frozen shoulder will improve over time, although it may take a long time up to year. The symptoms of frozen shoulder present by pain around shoulder and consequently limit range of motions. The effect of frozen shoulder leads to limit activities daily living life and high medical care cost. Physiotherapy is well known treatment for frozen shoulder but there was no data about the treatment of physiotherapy in frozen shoulder and length of follow up. Thus the aim of this study was to investigate physiotherapy program for frozen shoulder on range of motion and length of follow up. A retrospective study design was conducted. 469 medical records of patients with frozen shoulder were reviewed. These frozen shoulders were treated at physiotherapy unit, department of Rehabilitation last 3 years (January, 2014- December, 2016). The data consist of range of motions and length of follow up was recorded. The medical record of 183 males and 286 females with average aged 57.82±12.32 years were reviewed in this study. There was a statistically significant increase in shoulder flexion [mean difference 30.24 with 95%CI were [24.37-36.12], shoulder abduction [mean difference 34.93 with 95%CI were 27.8-42.0], shoulder internal rotation [mean difference 17.25 with 95%CI were 12.55-21.95] and shoulder external rotation [mean difference 17.71 with 95%CI were [13.07-22.36] respectively. In addition, the length of follow up averaged 84 days. In summary, the retrospective study show physiotherapy program likely to be benefit for patients with frozen shoulder in term of range of motion and short length of follow up.

Keywords: frozen shoulder, physiotherapy, range of motions, length of follow up

Procedia PDF Downloads 152

Authors: Hwang Kwon

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Objective: To assess the efficacy of embryo transfer (ET) of accumulated vitrified embryos and compare pregnancy outcomes between ET of thawed embryos following accumulation of vitrified embryos (frozen ET) and ET of fresh and thawed frozen embryos following accumulation of vitrified embryos (fresh ET + frozen ET). Study design: Patients were poor ovarian responders defined according to the Bologna criteria as well as a subgroup of women whose previous IVF-ET cycle through controlled ovarian stimulation (COS) yielded one or no embryos. Sixty-four frozen ETs were performed following accumulation of vitrified embryos (ACCE )(ACCE Frozen) and 51 fresh + frozen ETs were performed following accumulation of vitrified embryos (ACCE Fresh + Frozen). Positive βhCG rate, clinical pregnancy rate, ongoing pregnancy rate, and good quality embryos (%, ±SD) were compared between two groups. Results: There were more good quality embryos in the ACCE Fresh + Frozen group than in the ACCE Frozen group: 60±34.7 versus 42.9±28.9, respectively (p=0.03). Positive βhCG rate [18/64(28.2%) vs. 13/51(25.5%); p=0.75] and clinical pregnancy rate [12/64 (18.8%) vs. 11/51 (10.9%); p=0.71] were comparable between the two groups. Conclusion: Accumulation of vitrified embryos is an effective method in patients with poor ovarian response who fulfill the Bologna criteria. Pregnancy outcomes were comparable between the two groups.

Keywords: accumulation of embryos, frozen embryo transfer, poor responder, Bologna criteria

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Authors: Pınar Aydın, Şule Turhan

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Today, with the change of people's consumption habits, the importance of frozen fruit and vegetable sector has been increased. In Turkey, sector is based on export. It is growing very fast and external demand is constantly increasing. About 80% of frozen fruits and vegetables produced in Turkey are being exported. More than 90% of the exports go to European Union countries. About 49% of frozen fruits and vegetables in Turkey is being exported to Germany, England and France. In the sector which the abroad demand is continuously increasing, although it has been estimated that around 25% of the average annual growth rate, the domestic consumption is very low. Although the frozen food consumption per person in Turkey is about %2 of United States, the growing rate of the sector is higher than the United States and Europe. This situation reflects that it is such a sector that has a growing demand in both domestic and foreign markets.

Keywords: frozen food, fruit and vegetable sector, exports, Turkey

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Authors: Ayala Kobo-Greenhut, Amos Notea, Izhar Ben-Shlomo

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In R&D projects, there is no doubt about the need to change a current concept to an alternative one over time (i.e., concept leaping). Concept leaping is required since with most R&D projects uncertainty is present as they take place in dynamic environments. Despite the importance of concept leaping when needed, R&D teams may fail to do so (i.e., frozen concept). This research suggests a possible reason why frozen concept happens in the framework of quality engineering and control engineering. We suggest that frozen concept occurs since concept determines the derived plan and its implementation may be considered as equivalent to a closed-loop process, and is subject to the problem of not recognizing gaps as failures. We suggest that although implementing quality practices into an R&D project’s routine has many advantages, it intensifies the frozen concept problem since working according to quality practices relates to exploitation of learning behavior, while leaping to a new concept relates to exploring learning behavior.

Keywords: closed loop, control engineering, design, leaping, frozen concept, quality engineering, quality practices

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Authors: P. Thepnarintra

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Frozen shrimp industry plays an important role in the development of production industry of the country. There has been a continuing development to response the increasing demand; however, there have been some problems in running the enterprises. The purposes of this study are to: 1) investigate problems related to basic factors in operating frozen shrimp industry based on the entrepreneurs’ points of view. The enterprises involved in this study were small and medium industry receiving Thai Frozen Foods Association. 2) Compare the problems of the frozen shrimp industry according to their sizes of operation in 3 provinces of the central region Thailand. Population in this study consisted of 148 managers from 148 frozen shrimp enterprises Thai Frozen Foods Association, of which 77 were small size and 71 were medium size. The data were analyzed to find percentage, arithmetic mean, standard deviation, and independent sample T-test with the significant hypothesis at .05. The results revealed that the problems of the frozen shrimp industries of both size were in high level. The needs for government supporting were in high level. The comparison of the problems and the basic factors between the small and medium size enterprises showed no statistically significant level. The problems that they mentioned included raw materials, labors, production, marketing, and the need for academic supporting from the government sector.

Keywords: frozen shrimp industry, problems, related to the enterprise, operation

Procedia PDF Downloads 518

Authors: Narupon Kunbootsri, P. Sirasaporn

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Frozen shoulder is a common problem present by pain and limit range of motion. The prevalence of frozen shoulder showed 18-31% of population. The effect of frozen shoulder lead to limit activities daily living life, high medical care cost and so on. Physiotherapy is one of the treatments for frozen shoulder but there was no data about the treatment of physiotherapy. Moreover, it is question about onset of symptom relate to physiotherapy program and obtaining physical modalities and delayed start physiotherapy program lead to delayed improvement. Thus the aim of this study was to investigate physiotherapy program for frozen shoulder relate to onset of symptom, range of motion and obtaining physical modalities. A retrospective study design was conducted. 182 medical records of patients with frozen shoulder were reviewed. These frozen shoulders were treated at physiotherapy unit, department of Rehabilitation last 3 years (January, 2014- December, 2016). The data consist of onset of symptom, range of motion and obtaining physical modalities were recorded. There was a statistically significant increase in shoulder flexion [mean difference 38.88 with 95%CI were [16.00-61.77], shoulder abduction [mean difference 48.47 with 95%CI were 16.07-90.59], shoulder internal rotation [mean difference 22.36 with 95%CI were 2.81-37.18] and shoulder external rotation [mean difference 32.12 with 95%CI were [(-2.47)-(46.91)]. In addition, the onset of symptom was 76.42±46.90 days. And the physical modalities used frequently were hot pack 14.8% and ultrasound diathermy 13.7%. In conclusion, the physiotherapy program including, hot pack and ultrasound diathermy seem to be useful for frozen shoulder. But onset of symptom is too long to start physiotherapy programs.

Keywords: frozen shoulder, range of motions, onset of symptom, physiotherapy, physical modality

Procedia PDF Downloads 258

Authors: Narupon Kunbootsri, J. Kraipoj, K. Phandech, P. Sirasaporn

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Physiotherapy is one of the treatments for frozen shoulder but there was no data about the treatment of physiotherapy. Moreover, it is question about onset of symptom before physiotherapy program and obtaining physical modalities and delayed start physiotherapy program lead to delayed improvement. Thus the aim of this study was to investigate physiotherapy program for frozen shoulder on pain score, onset of symptom and obtaining physical modalities. A retrospective study design was conducted. 182 medical records of patients with frozen shoulder were reviewed. These frozen shoulders were treated at physiotherapy unit, department of Rehabilitation last 3 years (January, 2014- December, 2016). The data consist of onset of symptom, pain score and obtaining physical modalities were recorded. There was a statistically significant improve in pain score, pretreatment score mean 7.24±1.52 and the last follow up pain score mean 3.88± 1.0 [mean difference 3.18 with 95%CI were [2.45- 3.92]. In addition, the onset of symptoms was 145 days before obtaining physiotherapy program. The physical modalities used frequently were hot pack 14.8% and ultrasound diathermy 13.7%. In conclusion, the retrospective study show physiotherapy program including, hot pack and ultrasound diathermy seem to be useful for frozen shoulder in term of pain score. But onset of symptom is too long to start physiotherapy programs.

Keywords: frozen shoulder, physiotherapy, pain score, onset of symptom, physical modality

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Authors: R. Mehta, M. Ghogomu, B. Schoel

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Reports appeared in 2020 about China detecting SARS-Cov-2 (Covid-19) on frozen meat, shrimp, and food packaging material. In this study, we examined the use of swabs for the detection of Covid-19 on meat samples, and chicken breast (CB) was used as a model. Methods: Heat inactivated SARS-Cov-2 virus (IV) from Microbiologics was loaded onto the CB, swabbing was done, and the recovered inactivated virus was subjected to the Machery & Nagel NucleoSpin RNAVirus kit for RNA isolation according to manufacturer's instructions. For RT-PCR, the IDT 2019-nCoV RUO Covid-19 test kit was used with the Taqman Fast Virus 1-step master mix. The limit of detection (LOD) of viral load recovered from the CB was determined under various conditions: first on frozen CB where the IV was introduced on a defined area, then on frozen CB, with IV spread-out, and finally, on thawed CB. Results: The lowest amount of IV which can be reliably detected on frozen CB was a load of 1,000 - 2,000 IV copies where the IV was loaded on one spot of about 1 square inch. Next, the IV was spread out over a whole frozen CB about 16 square inches. The IV could be recovered at a lowest load of 4,000 to 8,000 copies. Furthermore, the effects of temperature change on viral load recovery was investigated i.e., if raw unfrozen meat became contaminated and remains for 1 hour at 4°C or gets refrozen. The amount of IV recovered successfully from CB kept at 4°C and the refrozen CB was similar to the recovery gotten from loading the IV directly on the frozen CB. In conclusion, an assay using swabs was successfully established for the detection of SARS-Cov-2 on frozen or raw (unfrozen) CB with a minimal load of up to 8,000 copies spread over 16 square inches.

Keywords: assay, COVID-19, meat, SARS-Cov-2

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Authors: Tramuta Clara, Masotti Chiara, Pitti Monica, Adriano Daniela, Battistini Roberta, Serraca Laura, Decastelli Lucia

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Oysters are considered filter organisms, and their raw consumption may increase health risks for consumers: it is often associated with outbreaks of gastroenteritis or enteric illnesses. Most of these foodborne diseases are caused by Vibrio strains, enteric pathogens also involved in the diffusion of genetic determinants of antibiotic resistance and their entrance along the food chain. The European Food Safety Authority (EFSA), during the European Union report on antimicrobial resistance in 2017, focused the attention about the role of food as a possible carrier of antibiotic-resistant bacteria or antibiotic-resistance genes that determine health risks for humans. This study wants to determine antibiotic resistance and antibiotic-resistance genes in Vibrio spp. isolated from Crassostrea gigas oysters collected in the Golfo della Spezia (Liguria, Italy). A total of 47 Vibrio spp. strains were isolated (ISO21872-2:2017) during the summer of 2021 from oysters of Crassostrea gigas. The strains were identified by MALDI-TOF (Bruker, Germany) mass spectrometry and tested for antibiotic susceptibility using a broth microdiluition method (ISO20776-1:2019) using Sensititre EUVSEC plates (Thermo-Fisher Scientific) to obtain the Minimum Inhibitory Concentration (MIC). The strains were tested with PCR-based biomolecular methods, according to previous works, to define the presence of 23 resistance genes of the main classes of antibiotics used in human and veterinary medicine: tet (B), tet (C), tet (D), tet (A), tet (E), tet (G ), tet (K), tet (L), tet (M), tet (O), tet (S) (tetracycline resistance); blaCTX-M, blaTEM, blaOXA, blaSHV (β-lactam resistance); mcr-1 and mcr-2 (colistin resistance); qnrA, qnrB, and qnrS (quinolone resistance); sul1, sul2 and sul3 (sulfonamide resistance). Six different species have been identified: V. alginolyticus 34% (n=16), V. harveyi 28% (n=13), V. fortis 15% (n=7), V. pelagius 8% (n=4), V. parahaemolyticus 11% (n=5) e V. chagasii 4% (n=2). The PCR assays showed the presence of the blaTEM gene on 40% of the strains (n=19). All the other genes were not detected, except for a V. alginolyticus positive for anrS gene. The broth microdiluition method results showed an high level of resistance for ciprofloxacin (62%; n=29), ampicillin (47%; n=22), and colistin (49%; n=23). Furthermore, 32% (n=15) of strains can be considered multiresistant bacteria for the simultaneous presence of resistance for three different antibiotic classes. Susceptibility towards meropenem, azithromycin, gentamicin, ceftazidime, cefotaxime, chloramphenicol, tetracycline and sulphamethoxazole reached 100%. The Vibrio species identified in this study are widespread in marine environments and can cause gastrointerstinal infections after the ingestion of raw fish products and bivalve molluscs. The level of resistance to antibiotics such as ampicillin, ciprofloxacin and colistin can be connected to anthropic factors (industrial, agricultural and domestic wastes) that promote the spread of resistance to these antibiotics. It can be also observed a strong correlation between phenotypic (resistant MIC) and genotypic (positive blaTEM gene) resistance for ampicillin on the same strains, probably due to the transfer of genetic material between bacterial strains. Consumption of raw bivalve molluscs can represent a risk for consumers heath due to the potentially presence of foodborne pathogens, highly resistant to different antibiotics and source of transferable antibiotic-resistant genes.

Keywords: vibrio species, blaTEM genes, antimicrobial resistance, PCR

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Authors: Nadia Miranda Lorick, Neela Badrie, Marsha Singh

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Cassava (Manihot esculenta, Crantz) which is also known as ‘yucca’ or ‘manioc’ has been acknowledged as a millennium crop which has been utilized for food security purposes. The crop provides considerable amount of energy. The aim of the study was to assess consumer groups of both ‘fresh’ and ‘frozen’ in terms of their perceptions toward the quality and safety attributes of frozen cassava. The questionnaire included four sections: consumer demographics, consumer perceptions on quality attributes of ‘frozen’ cassava, consumer knowledge, awareness and attitudes toward food safety of ‘frozen’ cassava and consumer suggestions toward the improvement of frozen cassava. A face-to-face questionnaire was administered to 200 consumers of cassava between April and May 2016. The criteria for inclusion in the survey were that they must be 15 years and over and consumer of cassava. The sections of the questionnaire included demographics of respondents, consumer perception on quality and safety attributes of cassava and suggestions for the improvement of the value-added product. The data was analysed by descriptive and chi-square using SPSS as well as qualitative information was captured. Only 17% of respondents purchased frozen cassava and this was significantly (P<0.05) associated to income. Some (15%) of fresh cassava purchasers had never heard of frozen cassava products and 7.5% o perceived that these products were unhealthy for consumption. More than half (51.3%) of the consumers (all from the ‘fresh’ cassava group) believed that there were ‘no toxins’ within cassava. The ‘frozen’ cassava products were valued for convenience but purchasers were least satisfied with ‘value for money’ (50%), ‘product safety’ (50%) and ‘colour’ (52.9%). Cassava purchasers demonstrated highest dissatisfaction levels with the quality attribute: value for money (6.6%, 11.8%) respectively. The most predominant area outlined by respondents for frozen cassava improvement was promotion /advertising/education (23%). The ‘frozen’ cassava purchasers were ‘least satisfied’ thus most concern that clean knives and clean surface would not be used agro- processing. Fresh cassava purchasers were comparatively more knowledgeable on the potential existence of naturally occurring toxins in cassava, however with 1% respondents being able to specifically identify the toxin as ‘cyanide’. Dangerous preservatives (31%), poor hygiene (30%) and chemicals from the packaging (11%) were identified as some sources of contamination of ‘frozen’ cassava. Purchasers of frozen cassava indicated that the information on packaging label was unclear (P<0.01) when compared to ‘fresh’ cassava consumers.

Keywords: consumer satisfaction, convenience, cyanide toxin, product safety, price, label

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Authors: Hyoun Joong Kim, Jin Woo Jun, Sib Sankar Giri, Cheng Chi, Saekil Yun, Sang Guen Kim, Sang Wha Kim, Jeong Woo Kang, Se Jin Han, Se Chang Park

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Vibrio corallilyticus is a well-known pathogen of coral. It is also infectious to a variety of shellfish species, including Pacific oyster (Crassostrea gigas) larvae. V. corallilyticus is remained to be a major constraint in marine bivalve aquaculture practice, especially in artificial seed production facility. Owing to the high mortality and contagious nature of the pathogen, large amount of antibiotics has been used for disease prevention and control. However, indiscriminate use of antibiotics may result in food and environmental pollution, and development of antibiotic resistant strains. Therefore, eco-friendly disease preventative measures are imperative for sustainable bivalve culture. The present investigation proposes the application of bacteriophage (phage) as an effective alternative method for controlling V. corallilyticus infection in marine bivalve hatcheries. Isolation of phages from sea water sample was carried out using drop or double layer agar methods. The host range, stability and morphology of the phage isolates were studied. In vivo phage efficacy to prevent V. corallilyticus infection in oyster larvae was also performed. The isolated phages, named pVco-5 and pVco-7 was classified as a podoviridae and pVco-14, was classified as a siphoviridae. Each phages were infective to four strains of seven V. corallilyticus strains tested. When oyster larvae were pre-treated with the phage before bacterial challenge, mortality of the treated oyster larvae was lower than that in the untreated control. This result suggests that each phages have the potential to be used as therapeutic agent for controlling V. corallilyticus infection in marine bivalve hatchery.

Keywords: bacteriophage, Vibrio coralliilyticus, Oyster larvae, mortality

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Authors: Abdurzag Kerban

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Preservation of semen had a major impact on sheep genetic breeding. The aim of this study was to evaluate the viability of ram spermatozoa after freezing pellet using cold surfaces made from cattle fat and paraffin wax. A pool of three to four ejaculates were pooled from six rams within a period of ten weeks. Semen was diluted in egg yolk-Tris diluent and processed in 0.25 ml straw and 0.1 ml pellets. Motility was evaluated after dilution, before freezing and post-thawing at 0, 1, 2 and 3 hour incubation. Viability index, acrosome integrity and leakage of intracellular enzymes (aspartat aminotransferase and alkline phosphatase) were also evaluated. Spermatozoa exhibited highly significant percentages of motility at 0, 1, 2 and 3 hours incubation after thawing and viability index in 0.25 ml straw and 0.1 ml pellets on cattle fat plate as compared to ram spermatozoa frozen on paraffin wax. In conclusion, cattle fat plate could be used as the cold surface of choice for freezing ram semen in form of pellets. Such form of frozen semen could be used as efficiently as semen frozen in straws. This simple method is economical with little expensive equipment or supplies, and may provide an efficient technique to cryopreserve ram spermatozoa in developing countries.

Keywords: ram semen, freezing, straw, pellet

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Authors: Nuralam Sam, Irawan Yusuf, Irfan Idris, Endi Adnan

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The most case in the shoulder in the the adult is the frozen shoulder. It make an uncomfortable sensation which disturbance daily activity. The studies of frozen shoulder are still limited. This study used a true experimental pre and post test design with a group design. The participant underwent dextrose prolotherapy injection in the rotator cuff, intraarticular glenohumeral joint, long head tendon biceps, and acromioclavicular joint injections with 15% dextrose, respectively, at week 2, week 4, and week 6. Participants were followed for 12 weeks. The specific biomarker MMP and TIMP, ROM, DASH score were measured at baseline, at week 6, and week 12. The data were analyzed by multivariate analysis (repeated measurement ANOVA, Paired T-Test, and Wilcoxon) to determine the effect of the intervention. The result showed a significant decrease in The Disability of the Arm, Shoulder, and Hand (DASH) score in prolo injection patients in each measurement week (p < 0.05). While the measurement of Range of Motion (ROM), each direction of shoulder motion showed a significant difference in average each week, from week 0 to week 6 (p <0.05).Dextrose prolotherapy injection results give a significant improvement in functional outcome of the shoulder joint, and ROMand did not show significant results in assessing the specific biomarker, MMP-1, and TIMP-1 in tissue repair. This study suggestion an alternative to the use of injection prolotherapy in Frozen shoulder patients, which has fewer side effects and better effectiveness than the use of corticosteroid injections.

Keywords: frozen shoulder, ROM, DASH score, prolotherapy, MMP-1, TIMP-1

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Authors: S. Amirhossein Tabatabaei, Abdulghader A. Aldaeef, Mohammad T. Rayhani

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As the permafrost continues to melt in the northern regions due to global warming, a soil-water mixture is left behind with drastically lower strength; a phenomenon that directly impacts the resilience of existing structures and infrastructure systems. The frozen soil-structure interaction, which in ice-poor soils is controlled by both interface shear and ice-bonding, changes its nature into a sole frictional state. Adfreeze, the controlling mechanism in frozen soil-structure interaction, diminishes as the ground temperature approaches zero. The main purpose of this paper is to capture the altered behaviour of frozen interface with respect to rising temperature, especially near melting states. A series of pull-out tests are conducted on model piles inside a cold room to study how the strength parameters are influenced by the phase change in ice-poor soils. Steel model piles, embedded in artificially frozen cohesionless soil, are subjected to both sustained pull-out forces and constant rates of displacement to observe the creep behaviour and acquire load-deformation curves, respectively. Temperature, as the main variable of interest, is increased from a lower limit of -10°C up to the point of melting. During different stages of the temperature rise, both skin deformations and temperatures are recorded at various depths along the pile shaft. Significant reduction of pullout capacity and accelerated creep behaviour is found to be the primary consequences of rising temperature. By investigating the different pull-out capacities and deformations measured during step-wise temperature change, characteristics of the transition from frozen to unfrozen soil-structure interaction are studied.

Keywords: Adfreeze, frozen soil-structure interface, ice-poor soils, pull-out capacity, warming permafrost

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Authors: Sayantika Biswas, Dipanshu Sur, Amitoj Athwal, Ratnabali Chakravorty

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Title: Impact of serum estrogen and progesterone levels in the outcome pregnancy rate in frozen embryo transfer cycles. A prospective cohort study Objective: The aim of the current study was to evaluate the effect of serum estradiol (E2) and progesterone (P4) levels at different time points on pregnancy outcomes in frozen embryo transfer (FET) cycles. Materials & Method: A prospective cohort study was performed in patients undergoing frozen embryo transfer. Patients under age 37 years of age with at least one good blastocyst or three good day 3 embryos were included in the study. For endometrial preparation, 14 days of oral estradiol use (2X2 mg for 5 days. 3X2 mg for 4 days, and 4X2 mg for 5 days) was followed by vaginal progesterone twice a day and 50 mg intramuscular progesterone twice a day. Embryo transfer was scheduled 72-76 hrs or 116-120hrs after the initiation of progesterone. Serum E2 and P4 levels were examined at 4 times a) at the start of the menstrual cycle prior to the hormone supplementation. b) on the day of P4 start. c) on the day of ET. d) on the third day after ET. Result: A total 41 women were included in this study (mean age 31.8; SD 2.8). Clinical pregnancy rate was 65.55%. Serum E2 levels on at the start of the menstrual cycle prior to the hormone supplementation and on the day of P4 start were high in patients who achieved pregnancy compared to who did not (P=0.005 and P=0.019 respectively). P4 levels on on the day of ET were also high in patients with clinical pregnancy. On the day of P4 start, a serum E2 threshold of 186.4 pg/ml had a sensitivity of 82%, and P4 had a sensitivity of 71% for the prediction of clinical pregnancy at the threshold value 16.00 ng/ml. Conclusion: In women undergoing FET with hormone replacement, serum E2 level >186.4 pg/ml on the day of the start of progesterone and serum P4 levels >16.00 ng/ml on embryo transfer day are associated with clinical pregnancy.

Keywords: serum estradiol, serum progesterone, clinical pregnancy, frozen embryo transfer

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Authors: Moussa I. Mohamed, Turki, K. A. Al-Faraj, Abdullah A. Al-Arfaj, Ashgan M. Hessain

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The objective of the present study is to detect the incidences of Salmonella enterica from different poultry farms located in Egypt on molecular basis. During the summer of 2012, a total of 1800 cloacal swabs were collected from poultry farms located I Cairo, Egypt to be subjected for isolation of Salmonella enteric. Moreover, a total of 300 samples of poultry and poultry products were collected from different retail establishment markets in Cairo, Egypt including, 150 local whole frozen chickens, 50 imported whole frozen chickens, 100 local chicken cut samples. The highest rate of isolation 8% was obtained from imported frozen chickens and local chicken cuts, followed by local frozen chickens 6.66% and finally rectal swabs from apparently health chickens 6.4 %. Salmonella Typhimurium and Salmonella Enteritidis were most frequent among the total Salmonella isolates. Multiplex-PCR for the rapid detection of Salmonella Typhimurium and Salmonella Enteritidis from field samples especially after pre-enrichment on Rappaport-Vassiliadis (RV) selective broth (PCR-RV), revealed the same positive samples. Therefore PCR-RV technique is rabid, time saving and applicable to detect Salmonella serovars directly from chicken samples. Moreover, detecting Salmonella Typhimurium and Salmonella Enteritidis by this assay was carried out within 2 days opposed to 5–6 d by the bacteriological and serological methods.

Keywords: Salmonella enterica, Salmonella typhimurium, Salmonella enteritidis enrichment, multiplex-PCR

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Authors: Bhavita S. Dave, Jaimin Vaidya, Chandresh H. Solanki, Atul K.

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To measure the permeability of a soil specimen, one of the basic assumptions of Darcy's law is that the soil sample should be saturated. Unlike saturated soils, the permeability of unsaturated soils cannot be found using conventional methods as it does not follow Darcy's law. Many empirical models, such as the Van Genuchten Model and Fredlund-Xing Model were suggested to predict permeability value for unsaturated soil. Such models use data from the soil-freezing characteristic curve to find fitting parameters for frozen unsaturated soils. In this study, soil specimens were subjected to 0, 1, 3, and 5 freezing-thawing (F-T) cycles for different degrees of saturation to have a wide range of suction, and its soil freezing characteristic curves were formulated for all F-T cycles. Changes in fitting parameters and relative permeability with subsequent F-T cycles are presented in this paper for both models.

Keywords: frozen unsaturated soil, Fredlund Xing model, soil-freezing characteristic curve, Van Genuchten model

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Authors: Muhammad Fiaz Qamar, Ahmad Faraz, Muhammad Arfan Zaman, Kazim Ali, Waleed Akram

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Babesiosis is reflected as the most important disease of cattle that are transmitted by arthropods. In Pakistan, its prevalence is up to 29% in the cattle and buffalo population in different regions. Cattle show a long lasting and durable immunity by giving an infection of B.bovis, B. bigemina, or Babesiadivergens. this is used in cattle to immunize them in a few countries as anti-babesiosis vaccine. Development of frozen vaccine allows for complete testing after production of each batch, However, once thawed, its reduced its shelf life, frozen vaccines are more difficult to transport as well as expensive to produce as compared to chilled vaccine. The contamination of blood derived vaccine has the potential risk that makes pre-production and post-production quality control necessary. For the trail master seed production of whole blood frozen bivalent Babesia(Babesiabovis and Babesiabigemina), 100 blood samples of Babesial positive suspected cattle was taken and processed for separation microscopic detection and rectification by PCR. Vaccine passages were done to reduce the parasitaemiasis in live calves. After 8 passages, parasitemia of Babesia reduced from 80% to 15%. Infected donor calf’s blood was taken by jugular cannulation by using preservative free lithium heparin as an anticoagulant (5 International Units IU heparin/ml blood). In lab, parasite containing blood was mixed in equal volumes with 3 M glycerol in PBS supplemented with 5 mM glucose (final concentration of glycerol 1.5 M) at 37°C. The mixture was then equilibrized at 37°C for 30 minutes and were dispensed in required containers (e.g., 5 ml cryovials).

Keywords: distribution, babesia, primer sequences, PCV

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Authors: Karina I. Hidas, Csaba Németh, Anna Visy, Judit Csonka, László Friedrich, Ildikó Cs. Nyulas-Zeke

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Egg yolk is a popular ingredient in the food industry due to its gelling, emulsifying, colouring, and coagulating properties. Because of the heat sensitivity of proteins, egg yolk can only be heat treated at low temperatures, so its shelf life, even with the addition of a preservative, is only a few weeks. Freezing can increase the shelf life of liquid egg yolk up to 1 year, but it undergoes gelling below -6 ° C, which is an irreversible phenomenon. The degree of gelation depends on the time and temperature of freezing and is influenced by the process of thawing. Therefore, in our experiment, we examined egg yolks thawed in different ways. In this study, unpasteurized, industrially broken, separated, and homogenized liquid egg yolk was used. Freshly produced samples were frozen in plastic containers at -18°C in a laboratory freezer. Frozen storage was performed for 90 days. Samples were analysed at day zero (unfrozen) and after frozen storage for 1, 7, 14, 30, 60 and 90 days. Samples were thawed in two ways (at 5°C for 24 hours and 30°C for 3 hours) before testing. Calorimetric properties were examined by differential scanning calorimetry, where heat flow curves were recorded. Denaturation enthalpy values were calculated by fitting a linear baseline, and denaturation temperature values were evaluated. Besides, dry matter content of samples was measured by the oven method with drying at 105°C to constant weight. For statistical analysis two-way ANOVA (α = 0.05) was employed, where thawing mode and freezing time were the fixed factors. Denaturation enthalpy values decreased from 1.1 to 0.47 at the end of the storage experiment, which represents a reduction of about 60%. The effect of freezing time was significant on these values, already the enthalpy of samples stored frozen for 1 day was significantly reduced. However, the mode of thawing did not significantly affect the denaturation enthalpy of the samples, and no interaction was seen between the two factors. The denaturation temperature and dry matter content did not change significantly either during the freezing period or during the defrosting mode. Results of our study show that slow freezing and frozen storage at -18°C greatly reduces the amount of protein that can be denatured in egg yolk, indicating that the proteins have been subjected to aggregation, denaturation or other protein conversions regardless of how they were thawed.

Keywords: denaturation enthalpy, differential scanning calorimetry, liquid egg yolk, slow freezing

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Authors: Adebisi M. Tiamiyu, Adewale F. Adeyemi, Olu-Ayobamikale V. Irewunmi

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Fish consumption has increased in recent years in both developing and advanced countries, owing to increased awareness of its nutritional and therapeutic benefits and its availability and affordability relative to other animal protein sources. Fish and fish products, however, are extremely prone to contamination by a wide range of hazardous organic and inorganic substances. This study assessed the levels of three heavy metals, copper (Cu), iron (Fe), and zinc (Zn), in frozen fish imported into Nigeria and sold in Ondo City for their safety for human consumption as recommended by WHO and FEPA. Three species of frozen fish (Scombrus scombrus, Merluccius merluccius, and Clupea harengus) were purchased, and the wet tissues (gills, muscles, and liver) were digested using a 3:1 mixture of nitric acid (HNO3) and hydrochloric acid (HCL). An atomic absorption spectrophotometer (AAS) was used to detect the amount of metal in the tissues. The levels of heavy metals in different fish species' organs varied. The fish had Zn > Fe > Cu heavy metal concentrations in that order. While the concentration of Cu and Fe in the tissues of all three fish species studied were within the WHO and FEPA prescribed limits for food fish, the concentration of Zn in the muscles of M. merluccius (0.262±0.052), C. harengus harengus (0.327±0.099), and S. scombrus (0.362±0.119) was above the prescribed limit (0.075 ppm) set by FEPA. An excessive amount of zinc in the body can cause nausea, headaches, decreased immunity, and appetite loss.

Keywords: heavy metal, atomic absorption spectrophotometer, fish, agencies

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Authors: Siti Rafeah Md Rafei, Karen Wang Yanping, Park Mi Kyoung

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Tissue samples are precious supply for molecular studies or disease identification diagnosed using molecular assays, namely real-time PCR (qPCR). It is critical to establish the most favorable nucleic acid extraction that gives the PCR-amplifiable genomic DNA. Furthermore, automated nucleic acid extraction is an appealing alternative to labor-intensive manual methods. Operational complexity, defined as the number of steps required to obtain an extracted sample, is one of the criteria in the comparison. Here we are comparing the One BioMed’s automated X8 platform with the commercially available manual-operated kits from QIAGEN Mini Kit and Roche. We extracted DNA from rat fresh-frozen tissue (from different type of organs) in the matrices. After tissue pre-treatment, it is added to the One BioMed’s X8 pre-filled cartridge, and the QIAGEN QIAmp column respectively. We found that the results after subjecting the eluates to the Real Time PCR using BIORAD CFX are comparable.

Keywords: DNA extraction, frozen tissue, PCR, qPCR, rat

Procedia PDF Downloads 122

Authors: Weishou Shen, Changxin Zou, Dong Liu

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High-mountain environments are experiencing more rapid warming than lowlands. The Tibetan (Qinghai-Xizang, TP) Plateau, known as the “Third Pole” of the Earth and the “Water Tower of Asia,” is the highest plateau in the world, however, ecological response to climate change has been hardly documented in high altitude regions. In this paper, we investigated climate warming induced ecological changes on the Tibetan Plateau over the past 50 years through combining remote sensing data with a large amount of in situ field observation. The results showed that climate warming up to 0.41 °C/10 a has greatly improved the heat conditions on the TP. Lake and river areas exhibit increased trend whereas swamp area decreased in the recent 35 years. The expansion in the area of the lake is directly related to the increase of precipitation as well as the climate warming up that makes the glacier shrink, the ice and snow melting water increase and the underground frozen soil melting water increase. Climate warming induced heat condition growth and reduced annual range of temperature, which will have a positive influence on vegetation, agriculture production and decreased freeze–thaw erosion on the TP. Terrestrial net primary production and farmland area on the TP have increased by 0.002 Pg C a⁻¹ and 46,000 ha, respectively. We also found that seasonal frozen soil depth decreased as the consequence of climate warming. In the long term, accelerated snow melting and thinned seasonal frozen soil induced by climate warming possibly will have a negative effect on alpine ecosystem stability and soil preservation.

Keywords: global warming, alpine ecosystem, ecological response, remote sensing

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Authors: Asma Ahmed Aljohani, Mohammed Orif

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The concentration of polycyclic aromatic hydrocarbons (PAH) in clam (A. glabrata) was examined in samples collected from Alseef Beach, 30 km south of Jeddah city. Gas chromatography-mass spectrometry (GC-MS) was used to analyse the 14 PAHs. The concentration of total PAHs was found to range from 11.521 to 40.149 ng/gdw with a mean concentration of 21.857 ng/gdw, which is lower compared to similar studies. The lower molecular weight PAHs with three rings comprised 18.14% of the total PAH concentrations in the clams, while the high molecular weight PAHs with four rings, five rings, and six rings account for 81.86%. Diagnostic ratios for PAH source distinction suggested pyrogenic or anthropogenic sources.

Keywords: bivalves, biomonitoring, hydrocarbons, PAHs

Procedia PDF Downloads 62

Authors: Sharon L. Gillies, Dario Jereb, Blayne Stam, Deji Arojojoye

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Plastic contamination of the oceans has become a global concern. Created during the breakdown of plastic or direct production, microplastic can be ingested by animals. This study aimed to assess the abundance of microplastic contamination in native salmon and bivalve species collected from the Salish Sea near the mouth of the Fraser River. Samples collected from May 2022 to September 2023 show that microplastic contamination was present in both salmon and bivalves. The most common were coloured microfibers, making up about 85% of the total microplastics, and the rest were uncoloured fibers and a few irregularly shaped microplastics.

Keywords: microplastics, microplastics in bivalves, microplastics in salmon, Salish Sea

Procedia PDF Downloads 49

Authors: Dilip Nath

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A study has been conducted on the available aquatic invertebrates in the river Dhansiri at Dimapur site. The study confirmed that the river body composed of aquatic macroinvertebrate community under two phyla viz., Arthropods and Molluscs. Total 10 species have been identified from there as the source of alternative protein food for the common people. Not only the protein source, they are also the component of aquatic food chain and indicators of aquatic ecosystem. Proper management and strategies to promote the edible invertebrates can be considered as the alternative protein and alternative income source for the common people for sustainable livelihood improvement.

Keywords: Dhansiri, Dimapur, invertebrates, livelihood improvement, protein

Procedia PDF Downloads 119

Authors: G. F. Okunade, M. O. Lawal, R. E. Uwadiae, D. Portnoy

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The diversity, physico-chemical, biochemical and genomics assessment of Macrofauna species of Ologe and Badagry Lagoons were carried out between August 2016 and July 2018. The concentrations of Fe, Zn, Mn, Cd, Cr, and Pb in water were determined by Atomic Absorption Spectrophotometer (AAS). Particle size distribution was determined with wet-sieving and sedimentation using hydrometer method. Genomics analyses were carried using 25 P. fusca (quadriseriata) and 25 P.fusca from each lagoon due to abundance in both lagoons all through the two years of collection. DNA was isolated from each sample using the Mag-Bind Blood and Tissue DNA HD 96 kit; a method designed to isolate high quality. The biochemical characteristics were analysed in the dominanat species (P.aurita and T. fuscatus) using ELISA kits. Physico-chemical parameters such as pH, total dissolved solids, dissolved oxygen, conductivity and TDS were analysed using APHA standard protocols. The Physico-chemical parameters of the water quality recorded with mean values of 32.46 ± 0.66mg/L and 41.93 ± 0.65 for COD, 27.28 ± 0.97 and 34.82 ± 0.1 mg/L for BOD, 0.04 ± 4.71 mg/L for DO, 6.65 and 6.58 for pH in Ologe and Badagry lagoons with significant variations (p ≤ 0.05) across seasons. The mean and standard deviation of salinity for Ologe and Badagry Lagoons ranged from 0.43 ± 0.30 to 0.27 ± 0.09. A total of 4210 species belonging to a phylum, two classes, four families and a total of 2008 species in Ologe lagoon while a phylum, two classes, 5 families and a total of 2202 species in Badagry lagoon. The percentage composition of the classes at Ologe lagoon had 99% gastropod and 1% bivalve, while Gastropod contributed 98.91% and bivalve 1.09% in Badagry lagoon. Particle size was distributed in 0.002mm to 2.00mm, particle size distribution in Ologe lagoon recorded 0.83% gravels, 97.83% sand, and 1.33% silt particles while Badagry lagoon recorded 7.43% sand, 24.71% silt, and 67.86% clay particles hence, the excessive dredging activities going on in the lagoon. Maximum percentage of sand (100%) was seen in station 6 in Ologe lagoon while the minimum (96%) was found in station 1. P. aurita (Ologe Lagoon) and T. fuscastus (Badagry Lagoon) were the most abundant benthic species in which both contributed 61.05% and 64.35%, respectively. The enzymatic activities of P. aurita observed with mean values of 21.03 mg/dl for AST, 10.33 mg/dl for ALP, 82.16 mg/dl for ALT and 73.06 mg/dl for CHO in Ologe Lagoon While T. fuscatus observed mean values of Badagry Lagoon) recorded mean values 29.76 mg/dl, ALP with 11.69mg/L, ALT with 140.58 mg/dl and CHO with 45.98 mg/dl. There were significant variations (P < 0.05) in AST and CHO levels of activities in the muscles of the species.

Keywords: benthos, biochemical responses, genomics, metals, particle size

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Authors: Nafees Ahmed, Nur Izyani Kamaruzman, Saralla Nathan, Mohd Ezharul Hoque Chowdhury, Anuar Zaini Md Zain, Iekhsan Othman, Sharifah Binti Syed Hassan

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Meat quality is always subject to consumer scrutiny when purchasing from retail markets on mislabeling as fresh meat. Various physiological and biochemical changes influence the quality of meat. As a major component of muscle tissue, proteins play a major role in muscle foods. In meat industry, freezing is the most common form of storage of meat products. Repeated cycles of freezing and thawing are common in restaurants, kitchen, and retail outlets and can also occur during transportation or storage. Temperature fluctuation is responsible for physical, chemical, and biochemical changes. Repeated cycles of ‘freeze-thaw’ degrade the quality of meat by stimulating the lipid oxidation and surface discoloration. The shelf life of meat is usually determined by its appearance, texture, color, flavor, microbial activity, and nutritive value and is influenced by frozen storage and subsequent thawing. The main deterioration of frozen meat during storage is due to protein. Due to the large price differences between fresh and frozen–thawed meat, it is of great interest to consumer to know whether a meat product is truly fresh or not. Researchers have mainly focused on the reduction of moisture loss due to freezing and thawing cycles of meat. The water holding capacity (WHC) of muscle proteins and reduced water content are key quality parameters of meat that ultimately changes color and texture. However, there has been limited progress towards understanding the actual mechanisms behind the meat quality changes under the freeze–thaw cycles. Furthermore, effect of freeze-thaw process on integrity of proteins is ignored. In this paper, we have studied the effect of ‘freeze-thawing’ on physicochemical changes of chicken meat protein. We have assessed the quality of meat by pH, spectroscopic measurements, Western Blot. Our results showed that increase in freeze-thaw cycles causes changes in pH. Measurements of absorbance (UV-visible and IR) indicated the degradation of proteins. The expression of various proteins (CREB, AKT, MAPK, GAPDH, and phosphorylated forms) were performed using Western Blot. These results indicated the repeated cycles of freeze-thaw is responsible for deterioration of protein, thus causing decrease in nutritious value of meat. It damges the use of these products in Islamic Sharia.

Keywords: chicken meat, freeze-thaw, halal, protein, western blot

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Authors: Eva Tvrdá, Marek Halenár, Hana Greifová, Alica Mackovich, Faridullah Hashim, Norbert Lukáč

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Oxidative stress associated with semen cryopreservation may result in lipid peroxidation (LPO), DNA damage and apoptosis, leading to decreased sperm motility and fertilization ability. Curcumin (CUR), a natural phenol isolated from Curcuma longa Linn. has been presented as a possible supplement for a more effective semen cryopreservation because of its antioxidant properties. This study focused to evaluate the effects of CUR on selected oxidative stress parameters in cryopreserved bovine semen. 20 bovine ejaculates were split into two aliquots and diluted with a commercial semen extender containing CUR (50 μmol/L) or no supplement (control), cooled to 4 °C, frozen and kept in liquid nitrogen. Frozen straws were thawed in a water bath for subsequent experiments. Computer assisted semen analysis was used to evaluate spermatozoa motility, and reactive oxygen species (ROS) generation was quantified by using luminometry. Superoxide generation was evaluated with the NBT test, and LPO was assessed via the TBARS assay. CUR supplementation significantly (P<0.001) increased the spermatozoa motility and provided a significantly higher protection against ROS (P<0.001) or superoxide (P<0.01) overgeneration caused by semen freezing and thawing. Furthermore, CUR administration resulted in a significantly (P<0.01) lower LPO of the experimental semen samples. In conclusion, CUR exhibits significant ROS-scavenging activities which may prevent oxidative insults to cryopreserved spermatozoa and thus may enhance the post-thaw functional activity of male gametes.

Keywords: bulls, cryopreservation, curcumin, lipid peroxidation, reactive oxygen species, spermatozoa

Procedia PDF Downloads 236

Authors: Lourdita Llanto

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This study described and assessed the implementation of artificial insemination (AI) for cattle and carabaos in the Bicol Region, Philippines: Albay, Sorsogon and Camarines Sur. Three hundred respondents were interviewed. Results were analyzed using frequency counts, means, percentages and chi-square test. Semen samples from different stations were analyzed for motility, viability and morphology. T-test was used in semen quality evaluation. Provincial AI coordinators (PAIC) were male, averaging 59 years old, married, had college education, served in government service for 34 years, but as PAIC for 5.7 years. All had other designations. Mean AI operation was 11.33 years with annual support from the local government unit of Php76,666.67. AI technicians were males, married, with college education, and trained on AI. Problems were on mobility; inadequate knowledge of farmers in animal raising and AI; and lack of liquid nitrogen and frozen semen supply. There was 2.95 municipalities and breedable cattle/carabaos of 3,091.25 per AI technician. Mean number of artificially inseminated animals per AI technician for 2011 was 28.57 heads for carabaos and 8.64 heads for cattle. There was very low participation rate among farmers. Carabaos were 6.52 years with parity 1.53. Cattle were 5.61 years, with parity of 1.51. Semen quality significantly (p ≤ 0.05) deteriorated in normal and live sperm with storage and handling at the provincial and field stations. Breed, AI technicians practices and AI operation significantly affected conception rate. Mean conception rate was 57.62%.

Keywords: artificial insemination, carabao, parity, mother tanks, frozen semen

Procedia PDF Downloads 416