Search results for: chicken embryos
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 285

Search results for: chicken embryos

45 The Impact of the Core Competencies in Business Management to the Existence and Progress of Traditional Foods Business with the Case of Study: Gudeg Sagan Yogyakarta

Authors: Lutfi AuliaRahman, Hari Rizki Ananda

Abstract:

The traditional food is a typical food of a certain region that has a taste of its own unique and typically consumed by a society in certain areas, one of which is Gudeg, a regional specialties traditional food of Yogyakarta and Central Java which is made of young jackfruit cooked in coconut milk, edible with rice and served with thick coconut milk (areh), chicken, eggs, tofu and sambal goreng krecek. However, lately, the image of traditional food has declined among people, so with gudeg, which today's society, especially among young people, tend to prefer modern types of food such as fast food and some other foods that are popular. Moreover, traditional food usually only preferred by consumers of local communities and lack of demand by consumers from different areas for different tastes. Thus, the traditional food producers increasingly marginalized and their consumers are on the wane. This study aimed to evaluate the management used by producers of traditional food with a case study of Gudeg Sagan which located in the city of Yogyakarta, with the ability of their management in creating core competencies, which includes the competence of cost, competence of flexibility, competence of quality, competence of time, and value-based competence. And then, in addition to surviving and continuing to exist with the existing external environment, Gudeg Sagan can increase the number of consumers and also reach a broader segment of teenagers and adults as well as consumers from different areas. And finally, in this paper will be found positive impact on the creation of the core competencies of the existence and progress of the traditional food business based on case study of Gudeg Sagan.

Keywords: Gudeg Sagan, traditional food, core competencies, existence

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44 Further Evidence for the Existence of Broiler Chicken PFN (Pale, Firm and Non-Exudative Meat) and PSE (Pale, Soft and Exudative) in Brazilian Commercial Flocks

Authors: Leila M. Carvalho, Maria Erica S. Oliveira, Arnoud C. Neto, Elza I. Ida, Massami Shimokomaki, Marta S. Madruga

Abstract:

The quality of broiler breast meat is changing as a result of the continuing emphasis on genetic selection for a more efficient meat production. Breast meat has been classified as PSE (pale, soft, exudative), DFD (dark, firm, dry) and normal color meat, and recently a third group has emerged: the so-called PFN (pale, firm, non-exudative) meat. This classification was based on pH, color and functional properties. The aim of this work was to confirm the existence of PFN and PSE meat by biochemical characterization and functional properties. Twenty four hours of refrigerated fillet, Pectoralis major, m. samples (n= 838) were taken from Cobb flocks 42-48 days old, obtained in Northeastern Brazil tropical region, the Northeastern, considered to have only dry and wet seasons. Color (L*), pH, water holding capacity (WHC), values were evaluated and compared with PSE group samples. These samples were classified as Normal (465.8), PSE meat (L*≥53; pH<5.8) and PFN (L*≥53; pH>5.8). The occurrence of control meat, PSE and PFN was 69.09%, 11.10% and 19.81%, respectively. Samples from PFN presented 4.0-5.0% higher WHC in relation to PSE meat and similar to control group. These results are explained by the fact that PSE meat syndrome occurs because of higher protein denaturation as the consequence of a simultaneous lower pH values under warm carcass sooner after slaughtering impairing the myofibril proteins functional properties. Conversely, PFN samples follow normal glycolysis rate maintaining the normal proteins activities. In conclusion, the results reported herein confirm the existence of this emerging broiler meat group with similar properties as control group and it should be considered as normal breast meat group.

Keywords: broiler breast meat, funcional properties, PFN, PSE

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43 Evaluation of Egg Quality Parameters in the Isa Brown Line in Intensive Production Systems in the Ocaña Region, Norte de Santander

Authors: Meza-Quintero Myriam, Lobo Torrado Katty Andrea, Sanchez Picon Yesenia, Hurtado-Lugo Naudin

Abstract:

The objective of the study was to evaluate the internal and external quality of the egg in the three production housing systems: floor, cage, and grazing of laying birds of the Isa Brown line, in the laying period between weeks 35 to 41; 135 hens distributed in 3 treatments of 45 birds per repetition were used (the replicas were the seven weeks of the trial). The feeding treatment supplied in the floor and cage systems contained 114 g/bird/day; for the grazing system, 14 grams less concentrate was provided. Nine eggs were collected to be studied and analyzed in the animal nutrition laboratory (3 eggs per housing system). The random statistical model was implemented: for the statistical analysis of the data, the statistical software of IBM® Statistical Products and Services Solution (SPSS) version 2.3 was used. The evaluation and follow-up instruments were the vernier caliper for the measurement in millimeters, a YolkFan™16 from Roche DSM for the evaluation of the egg yolk pigmentation, a digital scale for the measurement in grams, a micrometer for the measurement in millimeters and evaluation in the laboratory using dry matter, ashes, and ethereal extract. The results suggested that equivalent to the size of the egg (0.04 ± 3.55) and the thickness of the shell (0.46 ± 3.55), where P-Value> 0.05 was obtained, weight albumen (0.18 ± 3.55), albumen height (0.38 ± 3.55), yolk weight (0.64 ± 3.55), yolk height (0.54 ± 3.55) and for yolk pigmentation (1.23 ± 3.55). It was concluded that the hens in the three production systems, floor, cage, and grazing, did not show significant statistical differences in the internal and external quality of the chicken in the parameters studied egg for the production system.

Keywords: biological, territories, genetic resource, egg

Procedia PDF Downloads 56
42 Food Bolus Obstruction: A Rural Hospital’s Experience

Authors: Davina Von Hagt, Genevieve Gibbons, Matt Henderson, Tom Bowles

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Purpose: Food bolus obstructions are common emergency surgical presentations, but there is no established management guideline in a rural setting. Intervention usually involves endoscopic removal after initial medical management has failed. Within a rural setting, this falls upon the general surgeon. There are varied endoscopic techniques that may be used. Methodology: A review of the past fifty cases of food bolus obstruction managed at Albany Health Campus was retrospectively reviewed to assess endoscopic findings and techniques. Operation notes, histopathology, imaging, and patient notes were reviewed. Results: 50 patients underwent gastroscopy for food bolus obstruction from August 2017 to March 2021. Ages ranged from 11 months to 95 years, with the majority of patients aged between 30-70 years. 88% of patients were male. Meat was the most common bolus (20% unspecified, 20% steak, 10% chicken, 6% lamb, 4% sausage, 2% pork). At endoscopy, 12% were found not to have a food bolus obstruction. Two patients were found to have oesophageal cancer, and four patients had a stricture and required dilatation. A variety of methods were used to relieve oesophageal obstruction ranging from pushing through to stomach (24 patients), using an overtube (10 patients), raptor (13 patients), and less common instruments such as Roth net, basket, guidewire, and pronged grasper. One patient had an unsuccessful endoscopic retrieval and required theatre for laparoscopic assisted removal with rendezvous endoscopic piecemeal removal via oesophagus and gastrostomy. Conclusion: Food bolus obstruction is a common emergency presentation. Within the rural setting, management requires innovation and teamwork within the safety of the local experience.

Keywords: food bolus obstruction, regional hospital, surgical management, innovative surgical treatment

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41 Over Expression of Mapk8ip3 Patient Variants in Zebrafish to Establish a Spectrum of Phenotypes in a Rare-Neurodevelopmental Disorder

Authors: Kinnsley Travis, Camerron M. Crowder

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Mapk8ip3 (Mitogen-Activated Protein Kinase 8 Interacting Protein 3) is a gene that codes for the JIP3 protein, which is a part of the JIP scaffolding protein family. This protein is involved in axonal vesicle transport, elongation and regeneration. Variants in the Mapk8ip3 gene are associated with a rare-genetic condition that results in a neurodevelopmental disorder that can cause a range of phenotypes including global developmental delay and intellectual disability. Currently, there are 18 known individuals diagnosed to have sequenced confirmed Mapk8ip3 genetic disorders. This project focuses on examining the impact of a subset of missense patient variants on the Jip3 protein function by overexpressing the mRNA of these variants in a zebrafish knockout model for Jip3. Plasmids containing cDNA with individual missense variants were reverse transcribed, purified, and injected into single-cell zebrafish embryos (Wild Type, Jip3 -/+, and Jip3 -/-). At 6-days post mRNA microinjection, morphological, behavioral, and microscopic phenotypes were examined in zebrafish larvae. Morphologically, we compared the size and shape of the zebrafish during their development over a 5-day period. Total locomotive activity was assessed using the Microtracker assay and patterns of movement over time were examined using the DanioVision assay. Lastly, we used confocal microscopy to examine sensory axons for swelling and shortened length, which are phenotypes observed in the loss-of-function knockout Jip3 zebrafish model. Using these assays during embryonic development, we determined the impact of various missense variants on Jip3 protein function, compared to knockout and wild-type zebrafish embryo models. Variants in the gene Mapk8ip3 cause rare-neurodevelopmental disorders due to an essential role in axonal vesicle transport, elongation and regeneration. A subset of missense variants was examined by overexpressing the mRNA of these variants in a Jip3 knock-out zebrafish. Morphological, behavioral, and microscopic phenotypes were examined in zebrafish larvae. Using these assays, the spectrum of disorders can be phenotypically determined and the impact of variant location can be compared to knockout and wild-type zebrafish embryo models.

Keywords: rare disease, neurodevelopmental disorders, mrna overexpression, zebrafish research

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40 Biosafety Study of Genetically Modified CEMB Sugarcane on Animals for Glyphosate Tolerance

Authors: Aminah Salim, Idrees Ahmed Nasir, Abdul Qayyum Rao, Muhammad Ali, Muhammad Sohail Anjum, Ayesha Hameed, Bushra Tabassum, Anwar Khan, Arfan Ali, Mariyam Zameer, Tayyab Husnain

Abstract:

Risk assessment of transgenic herbicide tolerant sugarcane having CEMB codon optimized cp4EPSPS gene was done in present study. Fifteen days old chicks taken from K&Ns Company were randomly assorted into four groups with eight chicks in each group namely control chicken group fed with commercial diet, non-transgenic group fed with non-experimental sugarcane and transgenic group fed with transgenic sugarcane with minimum and maximum level. Body weights, biochemical analysis for Urea, alkaline phosphatase, alanine transferase, aspartate transferase, creatinine and bilirubin determination and histological examination of chicks fed with four types of feed was taken at fifteen days interval and no significant difference was observed in body weight biochemical and histological studies of all four groups. Protein isolated from the serum sample was analyzed through dipstick and SDS-PAGE, showing the absence of transgene protein in the serum sample of control and experimental groups. Moreover the amplification of cp4EPSPS gene with gene specific primers of DNA isolated from chicks blood and also from commercial diet was done to determine the presence and mobility of any nucleotide fragment of the transgene in/from feed and no amplification was obtained in feed as well as in blood extracted DNA of any group. Also no mRNA expression of cp4EPSPS gene was obtained in any tissue of four groups of chicks. From the results it is clear that there is no deleterious or harmful effect of the CEMB codon optimized transgenic cp4EPSPS sugarcane on the chicks health.

Keywords: chicks, cp4EPSPS, glyphosate, sugarcane

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39 A Statistical Approach to Predict and Classify the Commercial Hatchability of Chickens Using Extrinsic Parameters of Breeders and Eggs

Authors: M. S. Wickramarachchi, L. S. Nawarathna, C. M. B. Dematawewa

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Hatchery performance is critical for the profitability of poultry breeder operations. Some extrinsic parameters of eggs and breeders cause to increase or decrease the hatchability. This study aims to identify the affecting extrinsic parameters on the commercial hatchability of local chicken's eggs and determine the most efficient classification model with a hatchability rate greater than 90%. In this study, seven extrinsic parameters were considered: egg weight, moisture loss, breeders age, number of fertilised eggs, shell width, shell length, and shell thickness. Multiple linear regression was performed to determine the most influencing variable on hatchability. First, the correlation between each parameter and hatchability were checked. Then a multiple regression model was developed, and the accuracy of the fitted model was evaluated. Linear Discriminant Analysis (LDA), Classification and Regression Trees (CART), k-Nearest Neighbors (kNN), Support Vector Machines (SVM) with a linear kernel, and Random Forest (RF) algorithms were applied to classify the hatchability. This grouping process was conducted using binary classification techniques. Hatchability was negatively correlated with egg weight, breeders' age, shell width, shell length, and positive correlations were identified with moisture loss, number of fertilised eggs, and shell thickness. Multiple linear regression models were more accurate than single linear models regarding the highest coefficient of determination (R²) with 94% and minimum AIC and BIC values. According to the classification results, RF, CART, and kNN had performed the highest accuracy values 0.99, 0.975, and 0.972, respectively, for the commercial hatchery process. Therefore, the RF is the most appropriate machine learning algorithm for classifying the breeder outcomes, which are economically profitable or not, in a commercial hatchery.

Keywords: classification models, egg weight, fertilised eggs, multiple linear regression

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38 Epoxomicin Affects Proliferating Neural Progenitor Cells of Rat

Authors: Bahaa Eldin A. Fouda, Khaled N. Yossef, Mohamed Elhosseny, Ahmed Lotfy, Mohamed Salama, Mohamed Sobh

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Developmental neurotoxicity (DNT) entails the toxic effects imparted by various chemicals on the brain during the early childhood period. As human brains are vulnerable during this period, various chemicals would have their maximum effects on brains during early childhood. Some toxicants have been confirmed to induce developmental toxic effects on CNS e.g. lead, however; most of the agents cannot be identified with certainty due the defective nature of predictive toxicology models used. A novel alternative method that can overcome most of the limitations of conventional techniques is the use of 3D neurospheres system. This in-vitro system can recapitulate most of the changes during the period of brain development making it an ideal model for predicting neurotoxic effects. In the present study, we verified the possible DNT of epoxomicin which is a naturally occurring selective proteasome inhibitor with anti-inflammatory activity. Rat neural progenitor cells were isolated from rat embryos (E14) extracted from placental tissue. The cortices were aseptically dissected out from the brains of the fetuses and the tissues were triturated by repeated passage through a fire-polished constricted Pasteur pipette. The dispersed tissues were allowed to settle for 3 min. The supernatant was, then, transferred to a fresh tube and centrifuged at 1,000 g for 5 min. The pellet was placed in Hank’s balanced salt solution cultured as free-floating neurospheres in proliferation medium. Two doses of epoxomicin (1µM and 10µM) were used in cultured neuropsheres for a period of 14 days. For proliferation analysis, spheres were cultured in proliferation medium. After 0, 4, 5, 11, and 14 days, sphere size was determined by software analyses. The diameter of each neurosphere was measured and exported to excel file further to statistical analysis. For viability analysis, trypsin-EDTA solution were added to neurospheres for 3 min to dissociate them into single cells suspension, then viability evaluated by the Trypan Blue exclusion test. Epoxomicin was found to affect proliferation and viability of neuropsheres, these effects were positively correlated to doses and progress of time. This study confirms the DNT effects of epoxomicin on 3D neurospheres model. The effects on proliferation suggest possible gross morphologic changes while the decrease in viability propose possible focal lesion on exposure to epoxomicin during early childhood.

Keywords: neural progentor cells, epoxomicin, neurosphere, medical and health sciences

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37 Diversity of Dermatophytes and Keratinophilic Fungi from Inernational Tourist Spots, City of Taj Mahal

Authors: Harison Masih, Jyotsna Kiran Peter, Sundara Singh, Geetha Singh

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The present investigation deals with diversity of dermatophytes and keratinophilic fungi from different tourist spots such as Agra Fort, Akbar tomb, It-Mat-Ud-Daulah, Mariam tomb, Radha Swami Bagh, and Taj Mahal of Agra City. These fungi are medically important which causes various infections and diseases in humans and animals. The main reservoir of these pathogens are the keratinous substances that increases due to birds and animal activities in the vicinity of monuments, where thousands (5413266) annual visitors from all over the world are visiting. The soil samples were subjected to isolate the pathogenic fungi through bait technique (buffalo skin, chicken feathers, human hair and goat tail hair). Baits were spread over the soil samples and incubated at room temperature for 30-35 days and pure culture isolates were maintained in SDA medium, stored at 4°C. Highest number of visitors were (3906453) from Taj Mahal, minimum 10785 at Mariam tomb annually, the total 271 isolates were encountered from soil samples out of these 18 genera and 38 species were found in different season. Highest incidence was 4.79% frequency shown by Chrysosporium keratinophilum while least 738% frequency occurrence by Trichophyton simii in soil samples. From the present study it was concluded that the incidence of pathogenic fungal isolates were the common in tourists soil that are etiological agents of superficial mycosis. Thus, both human and animal activity seemed to play an important role in occurrence and distribution of keratinophilic and related dermatophytes at various tourist places of Agra city.

Keywords: dermatophytic fungal diversity, bait technique, visitors at tourist spots, human and animal activities, soil samples

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36 Isolation and Molecular Detection of Marek’s Disease Virus from Outbreak Cases in Chicken in South Western Ethiopia

Authors: Abdela Bulbula

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Background: Marek’s disease virus is a devastating infection, causing high morbidity and mortality in chickens in Ethiopia. Methods: The current study was conducted from March to November, 2021 with the general objective of performing antemortem and postmortem, isolation, and molecular detection of Marek’s disease virus from outbreak cases in southwestern Ethiopia. Accordingly, based on outbreak information reported from the study sites namely, Bedelle, Yayo, and Bonga towns in southwestern Ethiopia, 50 sick chickens were sampled. The backyard and intensive farming systems of chickens were included in the sampling and priorities were given for chickens that showed clinical signs that are characteristics of Marek’s disease. Results: By clinical examinations, paralysis of legs and wings, gray eye, loss of weight, difficulty in breathing, and depression were recorded on all chickens sampled for this study and death of diseased chickens was observed. In addition, enlargement of the spleen and gross lesions of the liver and heart were recorded during postmortem examination. The death of infected chickens was observed in both vaccinated and non-vaccinated flocks. Out of 50 pooled feather follicle samples, Marek’s disease virus was isolated from 14/50 (28%) by cell culture method and out of six tissue samples, the virus was isolated from 5/6(83.30%). By Real time polymerization chain reaction technique, which was targeted to detect the Meq gene, Marek’s disease virus was detected from 18/50 feather follicles which accounts for 36% of sampled chickens. Conclusion: In general, the current study showed that the circulating Marek’s disease virus in southwestern Ethiopia was caused by the oncogenic Gallid herpesvirus-2 (Serotype-1). Further research on molecular characterization of revolving virus in current and other regions is recommended for effective control of the disease through vaccination.

Keywords: Ethioi, Marek's disease, isolation, molecular

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35 A Study of Growth Performance, Carcass Characteristic, Meat Quality and Association of Polymorphism in the ApoVLDL-II Gene with Fat Accumulation in the Female Broiler, Thai Native and Betong Chickens (KU Line)

Authors: C. Kridtayopas, W. Danvilai, P. Sopannarath, A. Kayan, W. Loongyai

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Both Betong chicken (KU Line) and Thai Native chickens were the high quality of the meat and low carcass fat compared to broiler chickens. The objective of this study was to determine the growth performance, carcass characteristic, meat quality and association of polymorphism in the ApoVLDL-II gene with fat accumulation in the female broiler, Thai Native and Betong (KU line) chickens at 4-14 weeks. The chickens were used and reared under the same environment and management (100 chicks per breed). The results showed that body weight (BW) of broiler chickens was significantly higher than Thai Native and Betong (KU line) chickens (P < 0.01) through all the experiment. At 4-8 weeks of age, feed conversion ratio (FCR) of broiler chickens was significantly better than Thai Native and Betong (KU line) chickens (P < 0.01), then increased at week 8-14. The percentage of breast, abdominal fat and subcutaneous fat of broiler chickens was significantly greater than Thai Native and Betong (KU line) chickens (P < 0.01). However, Thai Native chickens showed the highest percentage of liver (P < 0.01) when compared to other breeds. In addition, the percentage of wing of Thai Native and Betong (KU line) chickens were significantly (P < 0.01) higher than broiler chickens. Meat quality was also determined and found that, pH of breast meat left from slaughter 45 minutes (pH45) and 24 hours (pH24) of broiler was significantly higher than Thai Native and Betong (KU line) (P < 0.01) whereas the percentage of drip loss, thawing loss, cooking loss and shear force was not significantly different between breeds. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique was used to genotype the polymorphism in the ApoVLDL-II gene in the broiler, Thai Native and Betong (KU line) chickens. The results found that, the polymorphism in the ApoVLDL-II gene at VLDL6 loci was not associated with fat accumulation in those studied population.

Keywords: ApoVLDL-II gene, Betong (KU line) chickens, broiler chickens, carcass characteristic, growth performance, meat quality, Thai native chickens

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34 Development of a Stable RNAi-Based Biological Control for Sheep Blowfly Using Bentonite Polymer Technology

Authors: Yunjia Yang, Peng Li, Gordon Xu, Timothy Mahony, Bing Zhang, Neena Mitter, Karishma Mody

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Sheep flystrike is one of the most economically important diseases affecting the Australian sheep and wool industry (>356M/annually). Currently, control of Lucillia cuprina relies almost exclusively on chemicals controls and the parasite has developed resistance to nearly all control chemicals used in the past. It is therefore critical to develop an alternative solution for the sustainable control and management of flystrike. RNA interference (RNAi) technologies have been successfully explored in multiple animal industries for developing parasites controls. This research project aims to develop a RNAi based biological control for sheep blowfly. Double-stranded RNA (dsRNA) has already proven successful against viruses, fungi and insects. However, the environmental instability of dsRNA is a major bottleneck for successful RNAi. Bentonite polymer (BenPol) technology can overcome this problem, as it can be tuned for the controlled release of dsRNA in the gut challenging pH environment of the blowfly larvae, prolonging its exposure time to and uptake by target cells. To investigate the potential of BenPol technology for dsRNA delivery, four different BenPol carriers were tested for their dsRNA loading capabilities, and three of them were found to be capable of affording dsRNA stability under multiple temperatures (4°C, 22°C, 40°C, 55°C) in sheep serum. Based on stability results, dsRNA from potential targeted genes was loaded onto BenPol carriers and tested in larvae feeding assays, three genes resulting in knockdowns. Meanwhile, a primary blowfly embryo cell line (BFEC) derived from L. cuprina embryos was successfully established, aim for an effective insect cell model for testing RNAi efficacy for preliminary assessments and screening. The results of this study establish that the dsRNA is stable when loaded on BenPol particles, unlike naked dsRNA rapidly degraded in sheep serum. The stable nanoparticle delivery system offered by BenPol technology can protect and increase the inherent stability of dsRNA molecules at higher temperatures in a complex biological fluid like serum, providing promise for its future use in enhancing animal protection.

Keywords: flystrike, RNA interference, bentonite polymer technology, Lucillia cuprina

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33 Catered Lunch Suspected Outbreak in a Garment Factory, Sleman District, Yogyakarta, Indonesia, 2017

Authors: Rieski Prihastuti, Meliana Depo, Trisno A. Wibowo, Misinem

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On October 19, 2017, Yogyakarta Islamic Hospital reported 38 garment employees with nausea, vomiting, headache, abdominal pain, and diarrhea after they had lunch on October 18, 2017, to Sleman District Health Office. Objectives of this study were to ensure the outbreak and identify source and route of transmission. Case-control study was conducted to analyze food items that caused the outbreak. A case was defined as a person who got symptoms such as abdominal pain, diarrhea, nausea with/without vomiting, fever, and headache after they had lunch on October 18, 2017. Samples included leftover lunch box, vomit, tap water and drinking water had been sent to the laboratory. Data were analyzed descriptively as frequency table and analyzed by using chi-square in bivariate analysis. All of 196 garment employee was included in this study. The common symptoms of this outbreak were abdominal pain (84.4%), diarrhea (72.8%), nausea (61.6%), headache (52.8%), vomiting (12.8%), and fever (6.4%) with median incubation period 13 hours (range 1-34 hours). Highest attack rate and odds ratio was found in grilled chicken (Attack Rate 58,49%) with Odds Ratio 11,023 (Confidence Interval 95% 1.383 - 87.859; p value 0,005). Almost all samples showed mold, except drinking water. Based on its sign and symptoms, also incubation period, diarrheal Bacillus cereus and Clostridium perfringens were suspected to be the causative agent of the outbreak. Limitation of this study was improper sample handling and no sample of food handler and stools in the food caterer. Outbreak investigation training needed to be given to the hospital worker, and monitoring should be done to the food caterer to prevent another outbreak.

Keywords: disease outbreak, foodborne disease, food poisoning, outbreak

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32 Impact of Two Herbal Seeds Supplementation on Growth Performance and Some Biochemical Blood and Tissue Parameters of Broiler Chickens

Authors: Hamada A. Ahmed, Kadry M. Sadek, Ayman E. Taha

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The effects of basil and/or chamomile seed supplementation on the growth of Hubbard broiler chicks were evaluated. The antioxidant effects of these supplements were also assessed. One hundred and twenty 1-day-old broiler chicks were randomly divided into four equal groups. The control group (group 1) was fed a basal diet (BD) without supplementation. Groups 2, 3, and 4 were fed the BD supplemented with 10g basil, 10g chamomile, and 5g basil plus 5g chamomile per kg of food, respectively. Basil supplementation alone or in combination with chamomile non-significantly (P≥0.05) increased final body weight (3.2% and 0.3%, respectively) and weight gain (3.5% and 3.6%, respectively) over the experimental period. Chamomile supplementation alone non-significantly (P≥0.05) reduced final body weight and weight gain over the experimental period by 1.7% and 1.7%, respectively. In comparison to the control group, herbal seed supplementation reduced feed intake and improved the feed conversion and protein efficiency ratios. In general, basil seed supplementation stimulated chicken growth and improved the feed efficiency more effectively than chamomile seed supplementation. The antioxidant activities of basil and/or chamomile supplementation were examined in the thymus, bursa, and spleen. In chickens that received supplements, the level of malondialdehyde was significantly decreased, whereas the activities of glutathione, superoxide dismutase, and catalase were significantly increased (P<0.05). Supplementation of basil and/or chamomile did not affect blood protein levels, but had lipid-lowering effects as evidenced by reduced serum levels of total lipids, triglycerides, and cholesterol. In conclusion, supplementation of basil and/or chamomile improved growth parameters in broiler chicks and had antioxidant and blood lipid-lowering effects. These beneficial effects of basil and/or chamomile supplementation resulted in economically viable production of high-quality white meat containing no harmful residues.

Keywords: herbal additives, basil, chamomile, broiler, growth performance, antioxidant

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31 Expression of Inflammatory and Cell Death Genes and DNA Damage Induced by Endotoxic Shock in Laying Hens

Authors: Mariam G. Eshak, Ahmed Abbas, M. I. El-Sabry, M. M. Mashaly

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This investigation was conducted to determine the physiological response and evaluate the expression of inflammatory and cell death genes and DNA damage induced by endotoxic shock in laying hens. Endotoxic shock was induced by a single intravenous injection of 107 Escherichia coli (E. coli,) colony/hen. In the present study, 240 forty-week-old laying hens (H&N) were randomly assigned into 2 groups with 3 replicates of 40 birds each. Hens were reared in battery cages with wire floors in an open-sided housing system under natural conditions. Housing and general management practices were similar for all groups. At 42-wk of age, 45 hens from the first group (15 replicate) were infected with E. coli, while the same number of hens from the second group was injected with saline and served as a control. Heat shock protein-70 (HSP-70) expression, plasma corticosterone concentration, body temperature, and the gene expression of bax, caspase-3 activity, P38, Interlukin-1β (Il-1β), and tumor necrosis factor alpha (TNF-α) genes and DNA damage in the brain and liver were measured. Hens treated with E. coli showed significant (P≤0.05) increase of body temperature by 1.2 ᴼC and plasma corticosterone by 3 folds compared to the controls. Further, hens injected with E.Coli showed markedly over-expression of HSP-70 and increase DNA damage in brain and liver. These results were synchronized with activating cell death program since our data showed significant (P≤0.05) high expression of bax and caspase-3 activity genes in the brain and liver. These results were related to remarkable over-inflammation gene expression of P38, IL-1β, and TNF-α in brain and liver. In conclusion, our results indicate that endotoxic shock induces inflammatory physiological response and triggers cell death program by promoting P38, IL-1β, and TNF-α gene expression in the brain and liver.

Keywords: chicken, DNA damage, Escherichia coli, gene expression, inflammation

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30 Evaluation of Two DNA Extraction Methods for Minimal Porcine (Pork) Detection in Halal Food Sample Mixture Using Taqman Real-time PCR Technique

Authors: Duaa Mughal, Syeda Areeba Nadeem, Shakil Ahmed, Ishtiaq Ahmed Khan

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The identification of porcine DNA in Halal food items is critical to ensuring compliance with dietary restrictions and religious beliefs. In Islam, Porcine is prohibited as clearly mentioned in Quran (Surah Al-Baqrah, Ayat 173). The purpose of this study was to compare two DNA extraction procedures for detecting 0.001% of porcine DNA in processed Halal food sample mixtures containing chicken, camel, veal, turkey and goat meat using the TaqMan Real-Time PCR technology. In this research, two different commercial kit protocols were compared. The processed sample mixtures were prepared by spiking known concentration of porcine DNA to non-porcine food matrices. Afterwards, TaqMan Real-Time PCR technique was used to target a particular porcine gene from the extracted DNA samples, which was quantified after extraction. The results of the amplification were evaluated for sensitivity, specificity, and reproducibility. The results of the study demonstrated that two DNA extraction techniques can detect 0.01% of porcine DNA in mixture of Halal food samples. However, as compared to the alternative approach, Eurofins| GeneScan GeneSpin DNA Isolation kit showed more effective sensitivity and specificity. Furthermore, the commercial kit-based approach showed great repeatability with minimal variance across repeats. Quantification of DNA was done by using fluorometric assay. In conclusion, the comparison of DNA extraction methods for detecting porcine DNA in Halal food sample mixes using the TaqMan Real-Time PCR technology reveals that the commercial kit-based approach outperforms the other methods in terms of sensitivity, specificity, and repeatability. This research helps to promote the development of reliable and standardized techniques for detecting porcine DNA in Halal food items, religious conformity and assuring nutritional.

Keywords: real time PCR (qPCR), DNA extraction, porcine DNA, halal food authentication, religious conformity

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29 Facies Analysis and Depositional Environment of Late Cretaceous (Cenomanian) Lidam Formation, South East Sirt Basin, Libya

Authors: Miloud M. Abugares

Abstract:

This study concentrates on the facies analysis, cyclicity and depositional environment of the Upper Cretaceous (Cenomanian) carbonate ramp deposits of the Lidam Formation. Core description, petrographic analysis data from five wells in Hamid and 3V areas in the SE Sirt Basin, Libya were studied in detail. The Lidam Formation is one of the main oil producing carbonate reservoirs in Southeast Sirt Basin and this study represents one of the key detailed studies of this Formation. In this study, ten main facies have been identified. These facies are; Chicken-Wire Anhydrite Facies, Fine Replacive Dolomite Facies, Bioclastic Sandstone Facies, Laminated Shale Facies, Stromatolitic Laminated Mudstone Facies, Ostracod Bioturbated Wackestone Facies, Bioturbated Mollusc Packstone Facies, Foraminifera Bioclastic Packstone/Grainstone Facies Peloidal Ooidal Packstone/Grainstone Facies and Squamariacean/Coralline Algae Bindstone Facies. These deposits are inferred to have formed in supratidal sabkha, intertidal, semi-open restricted shallow lagoon and higher energy shallow shoal environments. The overall depositional setting is interpreted as have been deposited in inner carbonate ramp deposits. The best reservoir quality is encountered in Peloidal- Ooidal Packstone/Grainstone facies, these facies represents storm - dominated shoal to back shoal deposits and constitute the inner part of carbonate ramp deposits. The succession shows a conspicuous hierarchical cyclicity. Porous shoal and backshoal deposits form during maximum transgression system and early regression hemi-cycle of the Lidam Fm. However; oil producing from shoal and backshoal deposits which only occur in the upper intervals 15 - 20 feet, which forms the large scale transgressive cycle of the Upper Lidam Formation.

Keywords: Lidam Fm. Sirt Basin, Wackestone Facies, petrographic, intertidal

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28 Insect Cell-Based Models: Asutralian Sheep bBlowfly Lucilia Cuprina Embryo Primary Cell line Establishment and Transfection

Authors: Yunjia Yang, Peng Li, Gordon Xu, Timothy Mahony, Bing Zhang, Neena Mitter, Karishma Mody

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Sheep flystrike is one of the most economically important diseases affecting the Australian sheep and wool industry (>356M/annually). Currently, control of Lucillia cuprina relies almost exclusively on chemicals controls, and the parasite has developed resistance to nearly all control chemicals used in the past. It is, therefore, critical to develop an alternative solution for the sustainable control and management of flystrike. RNA interference (RNAi) technologies have been successfully explored in multiple animal industries for developing parasites controls. This research project aims to develop a RNAi based biological control for sheep blowfly. Double-stranded RNA (dsRNA) has already proven successful against viruses, fungi, and insects. However, the environmental instability of dsRNA is a major bottleneck for successful RNAi. Bentonite polymer (BenPol) technology can overcome this problem, as it can be tuned for the controlled release of dsRNA in the gut challenging pH environment of the blowfly larvae, prolonging its exposure time to and uptake by target cells. To investigate the potential of BenPol technology for dsRNA delivery, four different BenPol carriers were tested for their dsRNA loading capabilities, and three of them were found to be capable of affording dsRNA stability under multiple temperatures (4°C, 22°C, 40°C, 55°C) in sheep serum. Based on stability results, dsRNA from potential targeted genes was loaded onto BenPol carriers and tested in larvae feeding assays, three genes resulting in knockdowns. Meanwhile, a primary blowfly embryo cell line (BFEC) derived from L. cuprina embryos was successfully established, aim for an effective insect cell model for testing RNAi efficacy for preliminary assessments and screening. The results of this study establish that the dsRNA is stable when loaded on BenPol particles, unlike naked dsRNA rapidly degraded in sheep serum. The stable nanoparticle delivery system offered by BenPol technology can protect and increase the inherent stability of dsRNA molecules at higher temperatures in a complex biological fluid like serum, providing promise for its future use in enhancing animal protection.

Keywords: lucilia cuprina, primary cell line establishment, RNA interference, insect cell transfection

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27 Utilization of Oat in Rabbit Feed for the Development of Healthier Rabbit Meat and Its Impact on Human Blood Lipid Profile

Authors: Muhammad Rizwan Tariq, Muhammad Issa Khan, Zulfiqar Ahmad, Muhammad Adnan Nasir, Muhammad Sameem Javed, Sheraz Ahmed

Abstract:

Functional foods may be a good tool that can be simply utilized in reducing community health expenses. Regular consumption of rabbit meat can offer patrons with bioactive components because the manipulation in rabbit feed is much successful to raise the levels of conjugated linoleic acid, ecosapentaenoic acid, decosahexaenoic acid, polyunsaturated fatty acids, selenium, tocopherol etc. and to reduce the ω-3/ω-6 ratio which is performing a major role in curing of cardiovascular and several other diseases. In comparison to the meats of other species, rabbit meat has higher amounts of protein with essential amino acids, especially in the muscles and low cholesterol contents that also have elevated digestibility. The present study was carried out to develop the functional rabbit meat by modifying feed ingredient of rabbit diet. Thirty-day old rabbits were fed with feeds containing 2 % and 4 % oat. The feeding trial was carried out for eight weeks. Rabbits were divided into three different groups and reared for the period of two months. T0 rabbits were considered control group while T1 rabbits were reared on 4% oat, and T2 were on 2% oat in the feed. At the end of the 8 weeks, the rabbits were slaughtered. Results presented in this study concluded that 4 % oat seed supplementation enhanced n-3 PUFA in meat. It was observed that oat seed supplementation also reduced fat percentage in the meat. Utilization of oat in the feed of rabbits significantly affected the pH, protein, fat, textural and concentration of polyunsaturated fatty acids. A study trial was conducted in order to examine the impact of functional meat on the blood lipid profile of human subjects. They were given rabbit meat in comparison to the chicken meat for the period of one month. The cholesterol, triglycerides and low density lipoprotein were found to be lower in blood serum of human subject group treated with 4 % oat meat.

Keywords: functional food, functional rabbit meat, meat quality, rabbit

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26 Production Performance, Gut Microbial Count, Antibody Titer and Selected Welfare Indices of Broiler Birds Fed Higher Level of Animal Protein Concentrate With or Without Organic Acids Blend and Microencapsulated Phyto-Essential Oil

Authors: Ziaul Islam, Asad Sultan, Sarzamin Khan

Abstract:

Organic acids and micro encapsulated phyto essential oils have revealed great potential as an antibiotic replacement and as an additive to work tremendously for the health maintenance of broiler chicken. To explore more about organic acids, a total of 600 day-old broiler chicks (Cobb-500) were procured from a local hatchery and distributed into 5 treatment groups having 6 replicates of 20 birds each; the duration of the biological trial was of 35 days. Group T1 served as a control group that were fed on corn soy-based diet only. T2 were fed with a diet having 6% poultry by-product meal (PBM) diet, T3, T4, and T5 were served as the same diet as T2 but supplemented with an organic acid, phyto essential oils alone, and a combination, respectively. The findings declared significant improvement (p<0.05) in body weight gain and FCR in groups T3, T4, and T5 while feed intake was not affected. European broiler performance indicators like production efficiency factor (EPEF) and broiler index (EBI) were improved significantly (p<0.05) by the treatments T3, T4, and T5 compared with T1 and T2. Carcass evaluation depicted significantly better (p<0.05) dressed and eviscerated weight along with carcass yield (T3, T4, T5). Broilers fed organic acid and phyto essential oils supplemented diet had significantly lower (p<0.05) Clostridium perfringens, Escherichia coliand Salmonella and increased Lactobacillus counts. Likewise, antibody titer against ND, IB, and IBD were also significantly (p<0.05) improved by the treatments T3, T4 and T5compared with the T1and T2. Litter moisture content was significantly (p<0.05) reduced by treatmentsT3, T4, and T5 on day 28 and 35 compared with the T1 and T2. These findings of the present study revealed that supplementation of organic acids blend and phyto-essential oils as an as an substitute to improve the performance of broilers without the use of feed antibiotics in broilers fed with 6% poultry by-product meal based diet.

Keywords: organic acid, phyto essential oils, growth performance, PBM, gut health, microbiota, immunity

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25 Non Chemical-Based Natural Products in the Treatment and Control of Disease in Fish

Authors: Albert P. Ekanem, Austin I. Obiekezie, Elizabeth X. Ntia

Abstract:

Introduction: Some African plants and bile from animals have shown efficacies in the treatment and control of diseases in farmed fish. The background of the study is based on the fact the African rain forest is blessed with the abundance of medicinal plants that should be investigated for their use in the treatment of diseases. The significance of the study is informed by the fact that chemical-based substances accumulate in the tissues of food fish, thereby reducing the food values of such products and moreover, the continuous use of chemotherapeutics in the aquatic environments tends to degrade the affected environment. Methodology: Plants and animal products were extracted, purified and applied under in vitro and in vivo conditions to the affected organisms. Effective plants and bills were analyzed for biologically active substances responsible for the activities by both qualitative and HPLC methods. Results: Extracts of Carica papaya and Mucuna pruriens were effective in the treatment of Ichthyophthiriasis in goldfish (Carassius auratus auratus) with high host tolerance. Similarly, ectoparasitic monogeneans were effectively dislodged from the gills and skin of goldfish by the application of extracts of Piper guineense at therapeutic concentrations. Artemesia annua with known antimalarial activities in human was also effective against fish monogenean parasites of Clarias gariepinus in a concentration-related manner without detriments to the host. Effective antibacterial activities against Aeromonas and Pseudomonas diseases of the African catfish (Heterobranchus longifilis) were demonstrated in some plants such as Phylanthus amarus, Allium sativum, A. annua, and Citrus lemon. Bile from some animals (fish, goat, chicken, cow, and pig) showed great antibacterial activities against some gastrointestinal bacterial pathogens of fish. Conclusions: African plants and some animal bile have shown potential promise in the treatment of diseases in fish and other aquatic animals. The use of chemical-based substances for control of diseases in the aquatic environments should be restricted.

Keywords: control, diseases, fish, treatment

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24 Non Chemical-Based Natural Products in the Treatment and Control of Fish Diseases

Authors: Albert P. Ekanem, Austin I. Obiekezie, Elizabeth X. Ntia

Abstract:

Introduction: Some African plants and bile from animals have shown efficacies in the treatment and control of diseases in farmed fish. The background of the study is based on the fact the African rain forest is blessed with abundance of medicinal plants that should be investigated for their use in the treatment of diseases. The significance of the study is informed by the fact that chemical-based substances accumulates in the tissues of food fish, thereby reducing the food values of such products and moreover, the continuous use of chemotherapeutants in the aquatic environments tends to degrades the affected environment. Methodology: Plants and animal products were extracted, purified and applied under in vitro and in vivo conditions to the affected organisms. Effective plants and biles were analyzed for active biological substances responsible for the activities by both qualitative and HPLC methods. Results: Extracts of Carica papaya and Mucuna pruriens were effective in the treatment of Ichthyophthiriasis in goldfish (Carassius auratus auratus) with high host tolerance. Similarly, ectoparasitic monogeneans were effectively dislodged from the gills and skin of goldfish by the application of extracts of Piper guineense at therapeutic concentrations. Artemesia annua with known antimalarial activities in human was also effective against fish monogenean parasites of Clarias gariepinus in a concentration related manner without detriments to the host. Effective antibacterial activities against Aeromonas and Pseudomonas diseases of the African catfish (Heterobranchus longifilis) were demonstrated in some plants such as Phylanthus amarus, Allium sativum, A. annua, and Citrus lemon. Bile from some animals (fish, goat, chicken, cow, and pig) showed great antibacterial activities against some gastrointestinal bacterial pathogens of fish. Conclusions: African plants and some animal bile have shown potential promise in the treatment of diseases in fish and other aquatic animals. The use of chemical-based substances for control of diseases in the aquatic environments should be restricted.

Keywords: control, diseases, fish, natural products, treatment

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23 Hematological and Biochemical Indices of Starter Broiler Chickens Fed African Black Plum Seed Nut (Vitex Doniana) Meal

Authors: Obadire F. O., Obadire, S. O., Adeoti R. F., Pirgozliev V.

Abstract:

An experiment was conducted to determine the efficacy of utilizing African black plum seed nut (ABPNBD) meal on hematological and biochemical indices of broiler chicken ration formulated to substitute wheat offal. A total of 150- 1day-old, male Agrited birds were reared for 28 days of the experiment. The birds were assigned to five dietary treatments, with ten birds per treatment replicated 3 times. Experimental diets were formulated by supplementing the milled African black plum nut at (0, 5, 10, 12.5, and 15%) inclusion levels in the starter broiler’s ratio designated as T1 (control diet containing no ABPBD), Treatments (T2, 3,4 and 5) contained ABPNBD at 5, 10, 12.5, and 15%, respectively, in a completely randomized design. The hematological and biochemical indices of the birds were determined. The result revealed that all hematological parameters measured were significant (P <0.05) except for WBC. Increasing inclusion levels of ABPNBD decreased the PCV, HB, and RBC of the birds across the treatment groups. Birds fed 12.5 and 15% ABPNBD diets recorded the least of the parameters. The result of the serum biochemical indices showed significant (P < 0.05) influence for all parameters measured except for alanine transaminase (ALT), (AST), and creatinine. The total protein (TP), albumin, globulin, and glucose values were reduced across the treatment group as ABPNBD inclusion increased. Birds fed above 10% ABPNBD recorded the lowest value of TP, albumin, globulin, and glucose when compared with birds on a control diet and other treatments. The uric acid ranged from 3.85 to 2 .13 mmol/L, while creatinine ranged from 62.00 to 53.50 mmol/l. AST ranged between 8.50 u/l (5%) to 7.90 u/l (10%). ALT ranged between 7.50 u/l (12.5%) to 5.50 u/l (5 and 10%). In conclusion, dietary inclusion of African black plum up to 10% has no detrimental effect on the health of the starter chickens. Meanwhile, inclusion above 10% revealed a negative effect on some blood parameters measured. Therefore, African black plum should be supplemented with probable probiotics or subjected to different processing methods if to be used at a 15% inclusion level for optimal results.

Keywords: African black plum seed, starter broiler chickens, hematological and serum biochemical indices, (Vitex doniana)

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22 A Prospective Study on the Pattern of Antibiotics Use and Prevalence of Multidrug Resistant Escherichia Coli in Poultry Chickens and Its Correlation with Urinary Tract Infection

Authors: Stelvin Sebastian, Andriya Annie Tom, Joyalanna Babu, Merin Joshy

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Introduction: The worldwide increase in the use of antibiotics in poultry and livestock industry to treat and prevent bacterial diseases and as growth promoters in feeds has led to the problem of development of antibiotic resistance both in animals and human population. Aim: To study the pattern of antibiotic use and prevalence of multidrug-resistant Escherichia coli in poultry chickens in selected farms in Muvattupuzha and to compare the spread of multidrug-resistant bacteria from poultry environment to UTI patients. Methodology: Two farms from each of 6 localities in Muvattupuzha were selected. A questionnaire on the pattern of antibiotic use and various farming practices were surveyed from farms. From each farm, 60samples of fresh fecal matter, litter from inside, litter from the outside shed, agricultural soil and control soil were collected, and antimicrobial susceptibility testing of E. coli was done. Antibiogram of UTI patients was collected from the secondary care hospital included in the study, and those were compared with resistance patterns of poultry samples. Results: From survey response antibiotics such as ofloxacin, enrofloxacin, levofloxacin, ciprofloxacin, colistin, ceftriaxone, neomycin, cephalexin, and oxytetracycline were used for treatment and prevention of infections in poultry. 31of 48 samples (51.66%) showed E. coli growth. 7 of 15 antibiotics (46.6%) showed resistance. Ampicillin, amoxicillin, meropenem, tetracycline showed 100% resistance to all samples. Statistical analysis confirmed similar resistance pattern in the poultry environment and UTI patients for antibiotics such as ampicillin, amoxicillin, amikacin, and ofloxacin. Conclusion: E. coli were resistant not only to extended-spectrum beta-lactams but also to carbapenems, which may be disseminated to the environment where litter was used as manure. This may due to irrational use of antibiotics in chicken or from their use in poultry feed as growth promoters. The study concludes the presence of multidrug-resistant E.coli in poultry and its spread to environment and humans, which may cause potentially serious implications for human health.

Keywords: multidrug resistance, escherichia coli, urinary tract infection, poultry

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21 Surveillance of Mycoplasma gallisepticum in Pet, Game and Free Flying Birds

Authors: Shamas Ul Hassan, Nasir Mukhtar, Sajjad Ur Rehman, Asghar Ali Mian, Iftikhar Hussain, Muhammad Safdar Anjum

Abstract:

The Mycoplasma gallisepticum (MG) is the major cause of economic looses in birds which is transmitted by free flying birds in the environment. These demands for improving the biosecurity measures at farm level including proper disposal of farm mortality and other wastes along with the inclusion of zoos and wild life parks in the MG surveillance programme. For the purpose of doing surveillance of MG in different pet, game and free flying birds a total of 12 samples each of peacocks, pheasants, ducks, pigeons, parrots, and house crows were included in the first ever study of its nature in Pakistan. During the study, the relevant samples along with recording clinical and postmortem findings were subjected to sero-prevalence, culture isolation and PCR system. Further PCR being more sensitive proves to be a better epidemiological tool. Seropositive findings revealed in peacocks, pheasants, ducks, pigeons, parrots, and crows were 66.7%, 58.3%, 41.7%, 41.7%, 16.7% and 16.7% respectively with some free flying birds giving ambiguous reactions. Whereas in the same order the culture/isolation positive results were recorded as 25%, 16.7%, 8.3%, 16.7%, 16.7%, and 25%. The samples were further confirmed on the basis of 732 bp product in PCR system. High rate of prevalence of MG in the pet, game and free flying birds regardless to their clinical findings demands to improve the biosecurity measures at the farm level with the minimum interaction of these birds with commercial poultry. Further the proper and timely disposal of all sorts of carcasses contaminated litter and wasted feed in such ways that the free flying birds are denied of picking up at those wastages. Moreover, MG surveillance system including the advances diagnostic techniques in wildlife parks and zoos be devised with proper timely preventive and therapeutic measures. The study proves that a variety of birds other then chicken either with or without clinical exhibitions carry MG organism which could be the potential source of infection for commercial poultry. The routine surveillance will be done to reduce the economic losses in poultry production.

Keywords: epidemiology, Mycoplasma gallisepticum (MG), free flying birds, surveillance, PCR

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20 Functional Analysis of Variants Implicated in Hearing Loss in a Cohort from Argentina: From Molecular Diagnosis to Pre-Clinical Research

Authors: Paula I. Buonfiglio, Carlos David Bruque, Lucia Salatino, Vanesa Lotersztein, Sebastián Menazzi, Paola Plazas, Ana Belén Elgoyhen, Viviana Dalamón

Abstract:

Hearing loss (HL) is the most prevalent sensorineural disorder affecting about 10% of the global population, with more than half due to genetic causes. About 1 in 500-1000 newborns present congenital HL. Most of the patients are non-syndromic with an autosomal recessive mode of inheritance. To date, more than 100 genes are related to HL. Therefore, the Whole-exome sequencing (WES) technique has become a cost-effective alternative approach for molecular diagnosis. Nevertheless, new challenges arise from the detection of novel variants, in particular missense changes, which can lead to a spectrum of genotype-to-phenotype correlations, which is not always straightforward. In this work, we aimed to identify the genetic causes of HL in isolated and familial cases by designing a multistep approach to analyze target genes related to hearing impairment. Moreover, we performed in silico and in vivo analyses in order to further study the effect of some of the novel variants identified in the hair cell function using the zebrafish model. A total of 650 patients were studied by Sanger Sequencing and Gap-PCR in GJB2 and GJB6 genes, respectively, diagnosing 15.5% of sporadic cases and 36% of familial ones. Overall, 50 different sequence variants were detected. Fifty of the undiagnosed patients with moderate HL were tested for deletions in STRC gene by Multiplex ligation-dependent probe amplification technique (MLPA), leading to 6% of diagnosis. After this initial screening, 50 families were selected to be analyzed by WES, achieving diagnosis in 44% of them. Half of the identified variants were novel. A missense variant in MYO6 gene detected in a family with postlingual HL was selected to be further analyzed. A protein modeling with AlphaFold2 software was performed, proving its pathogenic effect. In order to functionally validate this novel variant, a knockdown phenotype rescue assay in zebrafish was carried out. Injection of wild-type MYO6 mRNA in embryos rescued the phenotype, whereas using the mutant MYO6 mRNA (carrying c.2782C>A variant) had no effect. These results strongly suggest the deleterious effect of this variant on the mobility of stereocilia in zebrafish neuromasts, and hence on the auditory system. In the present work, we demonstrated that our algorithm is suitable for the sequential multigenic approach to HL in our cohort. These results highlight the importance of a combined strategy in order to identify candidate variants as well as the in silico and in vivo studies to analyze and prove their pathogenicity and accomplish a better understanding of the mechanisms underlying the physiopathology of the hearing impairment.

Keywords: diagnosis, genetics, hearing loss, in silico analysis, in vivo analysis, WES, zebrafish

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19 Treatment of Premalignant Lesions: Curcumin a Promising Non-Surgical Option

Authors: Heba A. Hazzah, Ragwa M. Farid, Maha M. A. Nasra, Mennatallah Zakria, Magda A. El Massik, Ossama Y. Abdallah

Abstract:

Introduction: Curcumin (Cur) is a polyphenol derived from the herbal remedy and dietary spice turmeric. It possesses diverse anti-inflammatory and anti-cancer properties following oral or topical administration. The buccal delivery of curcumin can be useful for both systemic and local disease treatments such as gingivitis, periodontal diseases, oral carcinomas, and precancerous oral lesions. Despite of its high activity, it suffers a limited application due to its low oral bioavailability, poor aqueous solubility, and instability. Aim: Preparation and characterization of curcumin solid lipid nanoparticles with a high loading capacity into a mucoadhesive gel for buccal application. Methodology: Curcumin was formulated as nanoparticles using different lipids, namely Gelucire 39/01, Gelucire 50/13, Precirol, Compritol, and Polaxomer 407 as a surfactant. The SLN were dispersed in a mucoadhesive gel matrix to be applied to the buccal mucosa. All formulations were evaluated for their content, entrapment efficiency, particle size, in vitro drug dialysis, ex vivo mucoadhesion test, and ex vivo permeation study using chicken buccal mucosa. Clinical evaluation was conducted on 15 cases suffering oral erythroplakia and erosive lichen planus. Results: The results showed high entrapment efficiency reaching almost 90 % using Gelucire 50, the loaded gel with Cur-SLN showed good adhesion property and 25 minutes in vivo residence time. In addition to stability enhancement for the Cur powder. All formulae did not show any drug permeated however, a significant amount of Cur was retained within the mucosal tissue. Pain and lesion sizes were significantly reduced upon topical treatment. Complete healing was observed after 6 weeks of treatment. Conclusion: These results open a room for the pharmaceutical technology to optimize the use of this golden magical powder to get the best out of it. In addition, the lack of local anti-inflammatory compounds with reduced side effects intensifies the importance of studying natural products for this purpose.

Keywords: curcumin, erythroplakia, mucoadhesive, pain, solid lipid nanoparticles

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18 Multi-Residue Analysis (GC-ECD) of Some Organochlorine Pesticides in Commercial Broiler Meat Marketed in Shivamogga City, Karnataka State, India

Authors: L. V. Lokesha, Jagadeesh S. Sanganal, Yogesh S. Gowda, Shekhar, N. B. Shridhar, N. Prakash, Prashantkumar Waghe, H. D. Narayanaswamy, Girish V. Kumar

Abstract:

Organochlorine (OC) insecticides are among the most important organotoxins and make a large group of pesticides. Physicochemical properties of these toxins, especially their lipophilicity, facilitate the absorption and storage of these toxins in the meat thus possess public health threat to humans. The presence of these toxins in broiler meat can be a quantitative and qualitative index for the presence of these toxins in animal bodies, which is attributed to Waste water of irrigation after spraying the crops, contaminated animal feeds with pesticides, polluted air are the potential sources of residues in animal products. Fifty broiler meat samples were collected from different retail outlets of Bengaluru city, Karnataka state, in ice cold conditions and later stored under -20°C until analysis. All the samples were subjected to Gas Chromatograph attached to Electron Capture Detector(GC-ECD, VARIAN make) screening and quantification of OC pesticides viz; Alachlor, Aldrin, Alpha-BHC, Beta-BHC, Dieldrin, Delta-BHC, o,p-DDE, p,p-DDE, o,p-DDD, p,p-DDD, o,p-DDT, p,p-DDT, Endosulfan-I, Endosulfan-II, Endosulfan Sulphate and Lindane(all the standards were procured from Merck). Extraction was undertaken by blending fifty grams (g) of meat sample with 50g Sodium Sulphate anahydrous, 120 ml of n-hexane, 120 ml acetone for 15 mins, extract is washed with distilled water and sample moisture is dried by sodium sulphate anahydrous, partitioning is done with 25 ml petroleum ether, 10 ml acetonitrile and 15 ml n-hexane shake vigorously for two minutes, sample clean up was done with florosil column. The reconstituted samples (using n-hexane) (Merck chem) were injected to Gas Chromatograph–Electron Capture Detector(GC-ECD). The present study reveals that, among the fifty chicken samples subjected for analysis, 60% (15/50), 32% (8/50), 28% (7/50), 20% (5/50) and 16% (4/50) of samples contaminated with DDTs, Delta-BHC, Dieldrin, Aldrin and Alachlor respectively. DDT metabolites, Delta-BHC were the most frequently detected OC pesticides. The detected levels of the pesticides were below the levels of MRL(according to Export Council of India notification for fresh poultry meat).

Keywords: accuracy, gas chromatography, meat, pesticide, petroleum ether

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17 Single Cell Rna Sequencing Operating from Benchside to Bedside: An Interesting Entry into Translational Genomics

Authors: Leo Nnamdi Ozurumba-Dwight

Abstract:

Single-cell genomic analytical systems have proved to be a platform to isolate bulk cells into selected single cells for genomic, proteomic, and related metabolomic studies. This is enabling systematic investigations of the level of heterogeneity in a diverse and wide pool of cell populations. Single cell technologies, embracing techniques such as high parameter flow cytometry, single-cell sequencing, and high-resolution images are playing vital roles in these investigations on messenger ribonucleic acid (mRNA) molecules and related gene expressions in tracking the nature and course of disease conditions. This entails targeted molecular investigations on unit cells that help us understand cell behavoiur and expressions, which can be examined for their health implications on the health state of patients. One of the vital good sides of single-cell RNA sequencing (scRNA seq) is its probing capacity to detect deranged or abnormal cell populations present within homogenously perceived pooled cells, which would have evaded cursory screening on the pooled cell populations of biological samples obtained as part of diagnostic procedures. Despite conduction of just single-cell transcriptome analysis, scRNAseq now permits comparison of the transcriptome of the individual cells, which can be evaluated for gene expressional patterns that depict areas of heterogeneity with pharmaceutical drug discovery and clinical treatment applications. It is vital to strictly work through the tools of investigations from wet lab to bioinformatics and computational tooled analyses. In the precise steps for scRNAseq, it is critical to do thorough and effective isolation of viable single cells from the tissues of interest using dependable techniques (such as FACS) before proceeding to lysis, as this enhances the appropriate picking of quality mRNA molecules for subsequent sequencing (such as by the use of Polymerase Chain Reaction machine). Interestingly, scRNAseq can be deployed to analyze various types of biological samples such as embryos, nervous systems, tumour cells, stem cells, lymphocytes, and haematopoietic cells. In haematopoietic cells, it can be used to stratify acute myeloid leukemia patterns in patients, sorting them out into cohorts that enable re-modeling of treatment regimens based on stratified presentations. In immunotherapy, it can furnish specialist clinician-immunologist with tools to re-model treatment for each patient, an attribute of precision medicine. Finally, the good predictive attribute of scRNAseq can help reduce the cost of treatment for patients, thus attracting more patients who would have otherwise been discouraged from seeking quality clinical consultation help due to perceived high cost. This is a positive paradigm shift for patients’ attitudes primed towards seeking treatment.

Keywords: immunotherapy, transcriptome, re-modeling, mRNA, scRNA-seq

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16 Physiological Indicators and Stress Index of Scavenging Chickens at Lafarge and Dangote Cement Factory Areas of Ogun State

Authors: Oluwadele Joshua Femi, Akinlabi Ebenezer Yemi, Onaopemipo Adeitan, Kazeem Bello, Anthony Ekeocha, Miraim Tawose

Abstract:

This study was carried out to determine the physiological and stress index of scavenging chickens in LAFARGE (Ewekoro) and Dangote (Ibese) Cement Factories Area of Ogun State. One hundred adult scavenging chickens comprising of 25 chickens from LAFARGE, Dangote and respective adjourning communities (Imasayi and Wasimi) were used. Experimental birds were caught at night on their perch and kept in cages till the next morning. Data were collected on rectal temperature, pulse rate, and respiratory rate of the birds. Also, 5ml blood was collected through the wing vein of the chickens in each location using a sterilized needle and syringe and transported to laboratory for analysis. Significant (P<0.05) highest pulse rate (215.64 beat/minute) and respiratory rate (19.90 breaths/minute) were recorded among scavenging chickens at LAFARGE (Ewekoro) Area and the least (198.61 beat/minute and 16.93 breaths/minute, respectively) at Imasayi. There was no significant (P>0.05) difference in the rectal temperature of the birds in the study area. Significant (P<0.05) differences were also recorded in the Packed Cell Volume (PCV), Hemoglobin (Hb), White Blood Cell (WBC), Monocyte, and Glucose level of the chickens in study area with the highest (P<0.05) Packed Cell Volume (28.06%) and Haemoglobin (4.01g/dl) recorded in Ibese and the least Packed Cell Volume (22.00%) and Haemoglobin (288g/dl) in Imasayi. Highest (P<0.05) Monocyte (4.28%) and glucose (256.53g/dl) were recorded among scavenging chickens at Dangote (Ibese) while the least Monocyte (0.00%) and Glucose (194.53g/dl) was recorded among chickens at Wasimi. Highest (P<0.05) White Blood Cell (6488.89×103µl) was recorded among chickens at Ewekoro and the lowest value in Ibese (4388.44×103µl). There was no significant (P>0.05) difference in the Heterophyl, Lymphocyte, Basophyl and Heterophyl/Lymphocyte ratio of the chickens in the study Area. The study concluded that chickens reared at LAFARGE (Ewekoro) were stressed and had comprised welfare and health status compared to Dangote (Ibese) cement area and other agrarian communities. Effective environmental mitigation programme should be put in place to enhance the welfare of the scavenging chickens in LAFARGE Cement Factory Area.

Keywords: blood, chicken, poisonous substances, pack cell volume, communities

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