Search results for: bovine rotavirus
238 Determination of Some Etiologic Agents in Calves with Diarrhea
Authors: Nermin Isik, Ozlem Derinbay Ekici, Oguzhan Avci
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The aim of this study was to determination of role infection in neonatal calves in Central Anatolian, Turkey. A total 300 fecal samples were collected from diarrheic neonatal calves, aged between 0–90 days from Konya, Karaman, and Aksaray from January to April 2014. Fecal specimens from calves with clinically diarrheic symptoms were examined for the presence of Bovine Coronavirus, Bovine Rotavirus, Cryptosporidium sp., and E. coli by commercially available capture direct enzyme linked immunosorbent assay (ELISA) kit and Modified Ziehl Neelsen method (MZN). Calves were grouped according to their age as follows: 1-14, 15-29, and 30-90 days. Cryptosporidium sp. infection was detected in 52.8%, 58.8%, and 39.2% by ELISA and 33.9%, 47%, 26.7% by MZN in the respective age groups. The seroprevalance of Rotavirus (12.5 %, 40 %, 12.5 %), Coronavirus (2.5%, 0%, 3.5%) and E. coli (5%, 4.7%, 8.9%) infections were determined according to the age groups respectively. Cryptosporidium sp. was the most detected enteropathogen (52 %) of calves and coronavirus was the least detected (2 %). The detection rate of the mixed enfection was 12.3%. In conclusion, it must be evaluated by mix infections in calves with diarrhea. These results will provide an important contribution against the factors that cause diarrheaKeywords: cryptosporidium sp., bovine coronavirus, bovine rotavirus, E.coli, calve, ELISA
Procedia PDF Downloads 541237 The Pathology of Bovine Rotavirus Infection in Calves That Confirmed by Enzyme Linked Immunosorbant Assay, Reverse Transcription Polymerase Chain Reaction and Real-Time RT-PCR
Authors: Shama Ranjan Barua, Tofazzal M. Rakib, Mohammad Alamgir Hossain, Tania Ferdushy, Sharmin Chowdhury
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Rotavirus is one of the main etiologies of neonatal diarrhea in bovine calves that causes significant economic loss in Bangladesh. The present study was carried out to investigate the pathology of neonatal enteritis in calves due to bovine rotavirus infection in south-eastern part of Bangladesh. Rotavirus was identified by using ELISA, RT-PCR (Reverse Transcription Polymerase Chain Reaction), real-time RT-PCR. We examined 12 dead calves with history of diarrhea during necropsy. Among 12 dead calves, in gross examination, 6 were found with pathological changes in intestine, 5 calves had congestion of small intestine and rest one had no distinct pathological changes. Intestinal contents and/or faecal samples of all dead calves were collected and examined to confirm the presence of bovine rotavirus A using Enzyme linked immunosorbant assay (ELISA), RT-PCR and real-time RT-PCR. Out 12 samples, 5 (42%) samples revealed presence of bovine rotavirus A in three diagnostic tests. The histopathological changes were found almost exclusively limited in the small intestine. The lesions of rotaviral enteritis ranged from slight to moderate shortening (atrophy) of villi in the jejunum and ileum with necrotic crypts. The villi were blunt and covered by immature epithelial cells. Infected cells, stained with Haematoxylin and Eosin staining method, showed characteristic syncytia and eosinophilc intracytoplasmic inclusion body. The presence of intracytoplasmic inclusion bodies in enterocytes is the indication of viral etiology. The presence of rotavirus in the affected tissues and/or lesions was confirmed by three different immunological and molecular tests. The findings of histopathological changes will be helpful in future diagnosis of rotaviral infection in dead calves.Keywords: calves, diarrhea, pathology, rotavirus
Procedia PDF Downloads 244236 Molecular Epidemiology of Rotavirus in Post-Vaccination Era in Pediatric Patients with Acute Gastroenteritis in Thailand
Authors: Nutthawadee Jampanil, Kattareeya Kumthip, Niwat Maneekarn, Pattara Khamrin
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Rotavirus A is one of the leading causes of acute gastroenteritis in children younger than five years of age, especially in low-income countries in Africa and South Asia. Two live-attenuated oral rotavirus vaccines, Rotarix and RotaTeq, have been introduced into routine immunization programs in many countries and have proven highly effective in reducing the burden of rotavirus-associated morbidity and mortality. In Thailand, Rotarix and RotaTeq vaccines have been included in the national childhood immunization program since 2020. The objectives of this research are to conduct a molecular epidemiological study and to characterize rotavirus genotypes circulating in pediatric patients with acute diarrhea in Chiang Mai, Thailand, from 2020-2022 after the implementation of rotavirus vaccines. Out of 858 stool specimens, 26 (3.0%) were positive for rotavirus A. G3P[8] (23.0%) was detected as the most predominant genotype, followed by G1P[8] (19.2%), G8P[8] (19.2%), G9P[8] (15.3%), G2P[4] (7.7%), G1P[6] (3.9%), G9P[4] (3.9%), and G8P[X] (3.9%). In addition, the uncommon rotavirus strain G3P[23] (3.9%) was also detected in this study, and this G3P[23] strain displayed a genetic background similar to the porcine rotavirus. In conclusion, there was a dramatic change in the prevalence of rotavirus A infection and the diversity of rotavirus A genotypes in pediatric patients in Chiang Mai, Northern Thailand, in the rotavirus post-vaccination period. The finding obtained from this research contributes to a better understanding of rotavirus epidemiology after rotavirus vaccine introduction. Furthermore, the identification of unusual G and P genotype combination strains provides significant evidence for the potential interspecies transmission between human and animal rotaviruses.Keywords: rotavirus, infectious disease, gastroenteritis, Thailand
Procedia PDF Downloads 56235 Diagnosis of Rotavirus Infection among Egyptian Children by Using Different Laboratory Techniques
Authors: Mohamed A. Alhammad, Hadia A. Abou-Donia, Mona H. Hashish, Mohamed N. Massoud
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Background: Rotavirus is the leading etiologic agent of severe diarrheal disease in infants and young children worldwide. The present study was aimed 1) to detect rotavirus infection as a cause of diarrhoea among children under 5 years of age using the two serological methods (ELISA and LA) and the PCR technique (2) to evaluate the three methodologies used for human RV detection in stool samples. Materials and Methods: This study was carried out on 247 children less than 5 years old, diagnosed clinically as acute gastroenteritis and attending Alexandria University Children Hospital at EL-Shatby. Rotavirus antigen was screened by ELISA and LA tests in all stool samples, whereas only 100 samples were subjected to RT-PCR method for detection of rotavirus RNA. Results: Out of the 247 studied cases with diarrhoea, rotavirus antigen was detected in 83 (33.6%) by ELISA and 73 (29.6%) by LA, while the 100 cases tested by RT-PCR showed that 44% of them had rotavirus RNA. Rotavirus diarrhoea was significantly presented with a marked seasonal peak during autumn and winter (61.4%). Conclusion: The present study confirms the huge burden of rotavirus as a major cause of acute diarrhoea in Egyptian infants and young children. It was concluded that; LA is equal in sensitivity to ELISA, ELISA is more specific than LA, and RT-PCR is more specific than ELISA and LA in diagnosis of rotavirus infection.Keywords: rotavirus, diarrhea, immunoenzyme techniques, latex fixation tests, RT-PCR
Procedia PDF Downloads 362234 Genotyping of Rotaviruses in Pediatric Patients with Gastroenteritis by Using Real-Time Reverse Transcription Polymerase Chain Reaction
Authors: Recep Kesli, Cengiz Demir, Riza Durmaz, Zekiye Bakkaloglu, Aysegul Bukulmez
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Objective: Acute diarrhea disease in children is a major cause of morbidity worldwide and is a leading cause of mortality, and it is the most common agent responsible for acute gastroenteritis in developing countries. With hospitalized children suffering from acute enteric disease up to 50% of the analyzed specimen were positive for rotavirus. Further molecular surveillance could provide a sound basis for improving the response to epidemic gastroenteritis and could provide data needed for the introduction of vaccination programmes in the country. The aim of this study was to investigate the prevalence of viral etiology of the gastroenteritis in children aged 0-6 years with acute gastroenteritis and to determine predominant genotypes of rotaviruses in the province of Afyonkarahisar, Turkey. Methods: An epidemiological study on rotavirus was carried out during 2016. Fecal samples obtained from the 144 rotavirus positive children with 0-6 years of ages and applied to the Pediatric Diseases Outpatient of ANS Research and Practice Hospital, Afyon Kocatepe University with the complaint of diarrhea. Bacterial agents causing gastroenteritis were excluded by using bacteriological culture methods and finally, no growth observed. Rotavirus antigen was examined by both the immunochromatographic (One Step Rotavirus and Adenovirus Combo Test, China) and ELISA (Premier Rotaclone, USA) methods in stool samples. Rotavirus RNA was detected by using one step real-time reverse transcription-polymerase chain reaction (RT-PCR). G and P genotypes were determined using RT-PCR with consensus primers of VP7 and VP4 genes, followed by semi nested type-specific multiplex PCR. Results: Of the total 144 rotavirus antigen-positive samples with RT-PCR, 4 (2,8%) were rejected, 95 (66%) were examined, and 45 (31,2%) have not been examined for PCR yet. Ninety-one (95,8%) of the 95 examined samples were found to be rotavirus positive with RT-PCR. Rotavirus subgenotyping distributions in G, P and G/P genotype groups were determined as; G1:45%, G2:27%, G3:13%, G9:13%, G4:1% and G12:1% for G genotype, and P[4]:33%, P[8]:66%, P[10]:1% for P genotype, and G1P[8]:%37, G2P[4]:%21, G3P[8]:%10, G4P[8]:%1, G9P[8]:%8, G2P[8]:%3 for G/P genotype . Not common genotype combination were %20 in G/P genotype. Conclusions: This study subscribes to the global agreement of the molecular epidemiology of rotavirus which will be useful in guiding the alternative and application of rotavirus vaccines or effective control and interception. Determining the diversity and rates of rotavirus genotypes will definitely provide guidelines for developing the most suitable vaccine.Keywords: gastroenteritis, genotyping, rotavirus, RT-PCR
Procedia PDF Downloads 232233 Genotyping of G/P No Typable Group a Rotavirus Strains Revealed G2 and G9 Genotype Circulations in Moroccan Children Fully Vaccinated with Rotarix™
Authors: H. Boulahyaoui, S. Alaoui Amine, C. Loutfi, H. El Annaz, N. Touil, El M. El Fahim, S. Mrani
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Three Moroccan children fully vaccinated with Rotarix™ have been hospitalized for Rotavirus Gastroenteritis (RVGE) in the pediatric division of the Farabi Hospital, Oujda. Rotavirus G/P genotypes could not be typed because of their delayed crossing threshold (Ct) resolute with a group A rotavirus (RVA) real time RT-PCR. These strains were adapted to cell culture. All viruses replicated and caused extensive cytopathic effects after four or five passages in MA104 cell lines. Significant improvements have been obtained in the amount of viral particles. Each virus multiplied to a high titer (7.5 TCID50/ml). VP7 and VP4 partial gene sequencing revealed distinct genotypes compared to the Rotarix(®) vaccine strain. Two strains were of G2P[4] genotype whereas the third was G9P[8] genotype. Virus isolation while labor intensive, is recommended as a second test, especially when higher sensitivity for conventional RVA genotyping RT-PCR is needed. VP7 antigenic similarities between these strains and Rotarix were determined.Keywords: esacpe-vaccine, Morocco, Rotarix, G2P[4], G9P[8]
Procedia PDF Downloads 322232 Sequence Analysis and Structural Implications of Rotavirus Capsid Proteins
Authors: Nishal Parbhoo, John B. Dewar, Samantha Gildenhuys
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Rotavirus is the major cause of severe gastroenteritis worldwide in children aged 5 and younger. Death rates are high particularly in developing countries. The mature rotavirus is a non-enveloped triple-layered nucleocapsid containing 11 double-stranded RNA segments. Here a global view on the sequence and structure of the three main capsid proteins, VP7, VP6, and VP2 is taken by generating a consensus sequence for each of these rotavirus proteins, for each species obtained from published data of representative rotavirus genotypes from across the world and across species. The degree of conservation between species was represented on homology models for each of the proteins. VP7 shows the highest level of variation with 14 - 45 amino acids showing conservation of less than 60%. These changes are localized to the outer surface which is exposed to antibodies alluding to a possible mechanism in evading the immune system. The middle layer, VP6 shows lower variability with only 14-32 sites having lower than 70% conservation. The inner structural layer made up of VP2 showed the lowest variability with only 1-16 sites having less than 70% conservation across species. The results correlate with proteins’ multiple structural roles. Although the nucleotide sequences vary due to an error-prone replication and lack of proofreading, the corresponding amino acid sequence of VP2, 6 and 7 remains conserved. Sequence conservation maintained for the virus results in stable protein structures, fit for function. This can be exploited in drug design, molecular studies and biotechnological applications.Keywords: amino acid sequence conservation, capsid protein, protein structure, vaccine candidate
Procedia PDF Downloads 287231 Efficiency on the Enteric Viral Removal in Four Potable Water Treatment Plants in Northeastern Colombia
Authors: Raquel Amanda Villamizar Gallardo, Oscar Orlando Ortíz Rodríguez
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Enteric viruses are cosmopolitan agents present in several environments including water. These viruses can cause different diseases including gastroenteritis, hepatitis, conjunctivitis, respiratory problems among others. Although in Colombia there are not regulations concerning to routine viral analysis of drinking water, an enhanced understanding of viral pollution and resistance to treatments is desired in order to assure pure water to the population. Viral detection is often complex due to the need of specialized and time-consuming procedures. In addition, viruses are highly diluted in water which is a drawback from the analytical point of view. To this end, a fast and selective detection method for detection enteric viruses (i.e. Hepatitis A and Rotavirus) were applied. Micro- magnetic particles were functionalized with monoclonal antibodies anti-Hepatitis and anti-Rotavirus and they were used to capture, concentrate and separate whole viral particles in raw water and drinking water samples from four treatment plants identified as CAR-01, MON-02, POR-03, TON-04 and located in the Northeastern Colombia. Viruses were molecularly by using RT-PCR One Step Superscript III. Each plant was analyzed at the entry and exit points, in order to determine the initial presence and eventual reduction of Hepatitis A and Rotavirus after disinfection. The results revealed the presence of both enteric viruses in a 100 % of raw water analyzed in all plants. This represents a potential health hazard, especially for those people whose use this water for agricultural purposes. However, in drinking water analysis, enteric viruses was only positive in CAR-01, where was found the presence of Rotavirus. As a conclusion, the results confirm Rotavirus as the best indicator to evaluate the efficacy of potable treatment plant in eliminating viruses. CAR potable water plant should improve their disinfection process in order to remove efficiently enteric viruses.Keywords: drinking water, hepatitis A, rotavirus, virus removal
Procedia PDF Downloads 222230 Enteropathogenic Viruses Associated with Acute Gastroenteritis among Under 5-Years Children in Africa: A Systematic Review and Meta-Analysis
Authors: Cornelius Arome Omatola, Ropo Ebenezer Ogunsakin, Anyebe Bernard Onoja, Martin-Luther Oseni Okolo, Joseph Abraham-Oyiguh, Kehinde Charles Mofolorunso, Phoebe Queen Akoh, Omebije Patience Adejo, Joshua Idakwo, Therisa Ojomideju Okeme, Danjuma Muhammed, David Moses Adaji, Sunday Ocholi Samson, Ruth Aminu, Monday Eneojo Akor
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Gastroenteritis viruses are the leading etiologic agents of diarrhea in children worldwide. We present data from thirty-three (33) eligible studies published between 2003 and 2023 from African countries bearing the brunt of the virus-associated diarrheal mortality. Random effects meta-analysis with proportion, subgroups, and meta-regression analyses were employed. Overall, rotavirus with estimated pooled prevalence of 31.0% (95% CI 24.0–39.0) predominated in all primary care visits and hospitalizations, followed by norovirus, adenovirus, sapovirus, astrovirus, and aichivirus with pooled prevalence estimated at 15.0% (95% CI 12.0–20.0), 10% (95% CI 6-15), 4.0% (95% CI 2.0–6.0), 4% (95% CI 3-6), and 2.3% (95% CI 1-3), respectively. Predominant rotavirus genotype was G1P[8] (38%), followed by G3P[8] (11.7%), G9P[8] (8.7%), and G2P[4] (7.1%); although, unusual genotypes were also observed, including G3P[6] (2.7%), G8P[6] (1.7%), G1P[6] (1.5%), G10P[8] (0.9%), G8P[4] (0.5%), and G4P[8] (0.4%). The genogroup II norovirus predominated over the genogroup I-associated infections (84.6%, 613/725 vs 14.9%, 108/725), with the GII.4 (79.3%) being the most prevalent circulating genotype. In conclusion, this review showed that rotavirus remains the leading driver of viral diarrhea requiring health care visits and hospitalization among under-five years children in Africa. Thus, improved rotavirus vaccination in the region and surveillance to determine the residual burden of rotavirus and the evolving trend of other enteric viruses are needed for effective control and management of cases.Keywords: enteric viruses, rotavirus, norovirus, adenovirus, astrovirus, gastroenteritis
Procedia PDF Downloads 77229 Molecular Detection of Acute Virus Infection in Children Hospitalized with Diarrhea in North India during 2014-2016
Authors: Ali Ilter Akdag, Pratima Ray
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Background:This acute gastroenteritis viruses such as rotavirus, astrovirus, and adenovirus are mainly responsible for diarrhea in children below < 5 years old. Molecular detection of these viruses is crucially important to the understand development of the effective cure. This study aimed to determine the prevalence of common these viruses in children < 5 years old presented with diarrhea from Lala Lajpat Rai Memorial Medical College (LLRM) centre (Meerut) North India, India Methods: Total 312 fecal samples were collected from diarrheal children duration 3 years: in year 2014 (n = 118), 2015 (n = 128) and 2016 (n = 66) ,< 5 years of age who presented with acute diarrhea at the Lala Lajpat Rai Memorial Medical College (LLRM) centre(Meerut) North India, India. All samples were the first detection by EIA/RT-PCR for rotaviruses, adenovirus and astrovirus. Results: In 312 samples from children with acute diarrhea in sample viral agent was found, rotavirus A was the most frequent virus identified (57 cases; 18.2%), followed by Astrovirus in 28 cases (8.9%), adenovirus in 21 cases (6.7%). Mixed infections were found in 14 cases, all of which presented with acute diarrhea (14/312; 4.48%). Conclusions: These viruses are a major cause of diarrhea in children <5 years old in North India. Rotavirus A is the most common etiological agent, follow by astrovirus. This surveillance is important to vaccine development of the entire population. There is variation detection of virus year wise due to differences in the season of sampling, method of sampling, hygiene condition, socioeconomic level of the entire people, enrolment criteria, and virus detection methods. It was found Astrovirus higher then Rotavirus in 2015, but overall three years study Rotavirus A is mainly responsible for causing severe diarrhea in children <5 years old in North India. It emphasizes the required for cost-effective diagnostic assays for Rotaviruses which would help to determine the disease burden.Keywords: adenovirus, Astrovirus, hospitalized children, Rotavirus
Procedia PDF Downloads 131228 Differential Antibrucella Activity of Bovine and Murine Macrophages
Authors: Raheela Akhtar, Zafar Iqbal Chaudhary, Yongqun Oliver He, Muhammad Younus, Aftab Ahmad Anjum
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Brucella abortus is an intracellular pathogen affecting macrophages. Macrophages release some components such as lysozymes (LZ), reactive oxygen species (ROS) and reactive nitrite intermediates (RNI) which are important tools against intracellular survival of Brucella. The antibrucella activity of bovine and murine macrophages was compared following stimulation with Brucella abortus lipopolysaccharides. Our results revealed that murine macrophages were ten times more potent to produce antibrucella components than bovine macrophages. The differential production of these components explained the differential Brucella killing ability of these species that was measured in terms of intramacrophagic survival of Brucella in murine and bovine macrophages.Keywords: bovine macrophages, Brucella abortus, cell stimulation, cytokines, Murine macrophages
Procedia PDF Downloads 551227 Multiple Etiologies and Incidences of Co-Infections in Childhood Diarrhea in a Hospital Based Screening Study in Odisha, India
Authors: Arpit K. Shrivastava, Nirmal K. Mohakud, Subrat Kumar, Priyadarshi S. Sahu
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Acute diarrhea is one of the major causes of morbidity and mortality among children less than five years of age. Multiple etiologies have been implicated for infectious gastroenteritis causing acute diarrhea. In our study fecal samples (n=165) were collected from children (<5 years) presenting with symptoms of acute diarrhea. Samples were screened for viral, bacterial, and parasitic etiologies such as Rotavirus, Adenovirus, Diarrhoeagenic Escherichia coli (EPEC, EHEC, STEC, O157, O111), Shigella spp., Salmonella spp., Vibrio cholera, Cryptosporidium spp., and Giardia spp. The overall results from our study showed that 57% of children below 5 years of age with acute diarrhea were positive for at least one infectious etiology. Diarrhoeagenic Escherichia coli was detected to be the major etiological agent (29.09%) followed by Rotavirus (24.24%), Shigella (21.21%), Adenovirus (5.45%), Cryptosporidium (2.42%), and Giardia (0.60%). Among the different DEC strains, EPEC was detected significantly higher in <2 years children in comparison to >2 years age group (p =0.001). Concurrent infections with two or more pathogens were observed in 47 of 160 (28.48%) cases with a predominant incidence particularly in <2-year-old children (66.66%) compared to children of 2 to 5 years age group. Co-infection of Rotavirus with Shigella was the most frequent combination, which was detected in 17.94% cases, followed by Rotavirus with EPEC (15.38%) and Shigella with STEC (12.82%). Detection of multiple infectious etiologies and diagnosis of the right causative agent(s) can immensely help in better management of acute childhood diarrhea. In future more studies focusing on the detection of cases with concurrent infections must be carried out, as we believe that the etiological agents might be complementing each other’s strategies of pathogenesis resulting in severe diarrhea.Keywords: children, co-infection, infectious diarrhea, Odisha
Procedia PDF Downloads 328226 Comparison of Real-Time PCR and FTIR with Chemometrics Technique in Analysing Halal Supplement Capsules
Authors: Mohd Sukri Hassan, Ahlam Inayatullah Badrul Munir, M. Husaini A. Rahman
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Halal authentication and verification in supplement capsules are highly required as the gelatine available in the market can be from halal or non-halal sources. It is an obligation for Muslim to consume and use the halal consumer goods. At present, real-time polymerase chain reaction (RT-PCR) is the most common technique being used for the detection of porcine and bovine DNA in gelatine due to high sensitivity of the technique and higher stability of DNA compared to protein. In this study, twenty samples of supplements capsules from different products with different Halal logos were analyzed for porcine and bovine DNA using RT-PCR. Standard bovine and porcine gelatine from eurofins at a range of concentration from 10-1 to 10-5 ng/µl were used to determine the linearity range, limit of detection and specificity on RT-PCR (SYBR Green method). RT-PCR detected porcine (two samples), bovine (four samples) and mixture of porcine and bovine (six samples). The samples were also tested using FT-IR technique where normalized peak of IR spectra were pre-processed using Savitsky Golay method before Principal Components Analysis (PCA) was performed on the database. Scores plot of PCA shows three clusters of samples; bovine, porcine and mixture (bovine and porcine). The RT-PCR and FT-IR with chemometrics technique were found to give same results for porcine gelatine samples which can be used for Halal authentication.Keywords: halal, real-time PCR, gelatine, chemometrics
Procedia PDF Downloads 228225 Sensitivity, Specificity and Efficiency Real-Time PCR Using SYBR Green Method to Determine Porcine and Bovine DNA Using Specific Primer Cytochrome B Gene
Authors: Ahlam Inayatullah Badrul Munir, M. Husaini A. Rahman, Mohd Sukri Hassan
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Real-time PCR is a molecular biology technique that is currently being widely used for halal services to differentiating between porcine and bovine DNA. The useful of technique become very important for student or workers (who works in the laboratory) to learn how the technique could be run smoothly without fail. Same concept with conventional PCR, real-time PCR also needed DNA template, primer, enzyme polymerase, dNTP, and buffer. The difference is in real-time PCR, have additional component namely fluorescent dye. The most common use of fluorescent dye in real-time PCR is SYBR green. The purpose of this study was to find out how sensitive, specific and efficient real-time PCR technique was combined with SYBR green method and specific primers of CYT b. The results showed that real-time PCR technique using SYBR Green, capable of detecting porcine and bovine DNA concentrations up to 0.0001 µl/ng. The level of efficiency for both types of DNA was 91% (90-110). Not only that in specific primer CYT b bovine primer could detect only bovine DNA, and porcine primer could detect only porcine primer. So, from the study could be concluded that real-time PCR technique that was combined with specific primer CYT b and SYBR green method, was sensitive, specific and efficient to detect porcine and bovine DNA.Keywords: sensitivity, specificity, efficiency, real-time PCR, SYBR green, Cytochrome b, porcine DNA, bovine DNA
Procedia PDF Downloads 305224 Evaluation of Disease Risk Variables in the Control of Bovine Tuberculosis
Authors: Berrin Şentürk
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In this study, due to the recurrence of bovine tuberculosis, in the same areas, the risk factors for the disease were determined and evaluated at the local level. This study was carried out in 32 farms where the disease was detected in the district and center of Samsun province in 2014. Predetermined risk factors, such as farm, environmental and economic risks, were investigated with the survey method. It was predetermined that risks in the three groups are similar to the risk variables of the disease on the global scale. These risk factors that increase the susceptibility of the infection must be understood by the herd owners. The risk-based contagious disease management system approach should be applied for bovine tuberculosis by farmers, animal health professionals and public and private sector decision makers.Keywords: bovine tuberculosis, disease management, control, outbreak, risk analysis
Procedia PDF Downloads 389223 PCR Detection, Histopathological Characterization, and Autogenous Immunization of Bovine Papillomatosis (Wart) in Cattle, in Mekelle, Northern Ethiopia
Authors: Kidane Workelul, Yohans Tekle, Guesh Negash, Haftay Abraha, Nigus Abebe Shumuye, Yisehak Tsegaye Redda
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Bovine papillomatosis (wart) is one of the economically important bovine skin diseases worldwide, caused by a group of viruses named papillomaviruses (PVs). However, it has often been misdiagnosed as other skin diseases and remained untreated. In order to determine the status of the diseases, twenty-two farms were visited, and fourteen infected cattle with cutaneous papillomatosis were identified from a total of 235. Papilloma biopsies were taken for molecular and histopathological characterization, the therapeutic trial of an autogenous vaccine was evaluated on infected animals. The overall status of bovine papillomatosis in this study was calculated as 5.96% (14/235). The disease was found to be statistically significant in the age groups less than two years (X² = 26.69, P = 0.0001). The more prominent histologically characterized lesions in the sampled tissue were identified as squamous papilloma and fibro-papilloma. The Polymerase Chain Reaction (PCR) based identification revealed that all the clinically and histo-pathologically characterized papillomatosis cases were found to be infected with Bovine Papilloma Virus1(BPV1), indicating that BPV1 was the most common and sole causative agent of the diseases in the study area. In immunizing active bovine papillomatosis, an autogenous vaccine therapeutic trial demonstrated excellent results, with practically full recovery and no recurrence of the infection. Hence, it is concluded that bovine papillomatosis is an economically important disease of young age group cattle as well as a treatable disease. So, the production of marketable autogenous vaccines against bovine papillomatosis should be started and given at an early stage.Keywords: autogenous vaccine, bovine papillomatosis, bovine papilloma virus1 clinical-pathology, polymerase chine reaction, wart
Procedia PDF Downloads 71222 Culturing of Bovine Pre-Compacted Morlae in TCM-199 and Baf in a Standard 5% CO2 Laboratory Incubator and in the Vagina of a Goat Doe
Authors: Daniel M. Barry
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Since more than half a century ago, attempts have been made to culture cells and embryos outside the body (in vitro or ex vivo). This was done with different culture media and in various “incubators”. In the present study two different culture media were used: a standard TCM-199 culture medium and first trimester amniotic fluid (BAF) collected sterilely from pregnant cows after slaughter. Two different culture conditions were also investigated, the standard laboratory CO2 incubator versus culturing bovine embryos in the vagina of a goat doe. Two experiments were done: Firstly the permeability of different receptacles to CO2 gas was analyzed for possible culture in the vagina. Four-well plates and straws were used to incubate TCM-199 and BAF for a period of 120 h in the presence or absence of 5% CO2 gas. The pH values were measured and recorded every 24 h. In the second experiment pre-compacted morula stage bovine embryos were cultured in the above culture media in sealed 0.25 mL straws in a standard laboratory incubator and in the vagina of a goat doe. Evaluation was done on (1) stage of development and (2) number of blastomeres after 96 h of culture. In the first experiment it was shown that the CO2 gas diffused out of the 4-well plate as well as through the wall of the straws in the absence of CO2 gas, while in the presence of CO2 the pH of both media stabilized between 7.3 and 7.5. This meant that the semen straws were permeable to CO2 gas and could therefore be used as receptacles for culturing early stage bovine embryos. In the second experiment no statistical differences (p>0.05) were found in the number of pre-compacted bovine embryos that developed to the blastocyst stage, or the hatched blastocyst stage, neither for the culture medium used, or the method of culturing in the two incubators. Neither was there any difference (p>0.05) in the number of blastomeres that developed at the blastocyst stage between the two types of incubators. The bovine embryos tended to develop more blastomeres when cultured in BAF than when cultured in TCM-199 in both the standard laboratory incubator and when using the vagina of a goat doe as an incubator.Keywords: alternative culture, bovine embryos, vagina, bovine amniotic fluid, incubator
Procedia PDF Downloads 481221 Adsorption of Bovine Serum Albumine on CeO2
Authors: Roman Marsalek
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Preparation of nano-particles of cerium oxide and adsorption of bovine serum albumine on them were studied. Particle size distribution and influence of pH on zeta potential of prepared CeO2 were determined. Average size of prepared cerium oxide nano-particles was 9 nm. The simultaneous measurements of the bovine serum albumine adsorption and zeta potential determination of the (adsorption) suspensions were carried out. The adsorption isotherms were found to be of typical Langmuir type; values of the bovine serum albumin adsorption capacities were calculated. Increasing of pH led to decrease of zeta potential and decrease of adsorption capacity of cerium oxide nano-particles. The maximum adsorption capacity was found for strongly acid suspension (am=118 mg/g). The samples of nanoceria with positive zeta potential adsorbed more bovine serum albumine on the other hand, the samples with negative zeta potential showed little or no protein adsorption. Surface charge or better say zeta potential of CeO2 nano-particles plays the key role in adsorption of proteins on such type of materials.Keywords: adsorption, BSA, cerium oxide nanoparticles, zeta potential, albumin
Procedia PDF Downloads 364220 Performance of High Density Genotyping in Sahiwal Cattle Breed
Authors: Hamid Mustafa, Huson J. Heather, Kim Eiusoo, Adeela Ajmal, Tad S. Sonstegard
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The objective of this study was to evaluate the informativeness of Bovine high density SNPs genotyping in Sahiwal cattle population. This is a first attempt to assess the Bovine HD SNP genotyping array in any Pakistani indigenous cattle population. To evaluate these SNPs on genome wide scale, we considered 777,962 SNPs spanning the whole autosomal and X chromosomes in Sahiwal cattle population. Fifteen (15) non related gDNA samples were genotyped with the bovine HD infinium. Approximately 500,939 SNPs were found polymorphic (MAF > 0.05) in Sahiwal cattle population. The results of this study indicate potential application of Bovine High Density SNP genotyping in Pakistani indigenous cattle population. The information generated from this array can be applied in genetic prediction, characterization and genome wide association studies of Pakistani Sahiwal cattle population.Keywords: Sahiwal cattle, polymorphic SNPs, genotyping, Pakistan
Procedia PDF Downloads 417219 Development and Evaluation of Novel Diagnostic Methods for Infectious Rhinotracheitis of Cattle
Authors: Wenxiao Liu, Kun Zhang, Yongqing Li
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Bovine herpesvirus 1, a member of the genus Variellovirus of the subfamily Alphaherpesvirinae, has caused severe economic cost to the bovine industry. In this study, BoHV-1 glycerol protein gD was expressed in insect cells, and the purified gD was immunized in the Balb/C mice to generate monoclonal antibodies. Based on hybridoma cell fusion techniques, 20 monoclonal antibodies against Bovine herpesvirus 1 have been obtained. Further, mAb 3F8 with neutralizing activity and gD were applied to develop a blocking enzyme-linked immunosorbent assay (Elisa) for detecting neutralizing antibodies against BoHV-1, which shows a significant correlation between the blocking Elisa and VNT. The sensitivity and specificity of the test were estimated to be 94.59% and 93.42%, respectively. Furthermore, antibody pairing tests revealed that mAb 1B6 conjugated to fluorescence microspheres was used as the capture antibody, and mAb 3F9 was used as the detectable antibody to establish the immunochromatographic assay (ICS). The ICS was conducted to detect BoHV-1 in bovine samples with high sensitivity, specificity, and good stability. Clinical sample testing revealed that the results of ICS and real-time PCR have a coincidence rate of 95.42%. Our research confirmed that the ICS is a rapid and reliable method for the diagnosis of BoHV-1. In conclusion, our results lay a solid foundation for the prevention and control of BoHV-1 infection.Keywords: bovine disease, BoHV-1, ELISA, ICS assay
Procedia PDF Downloads 61218 An Epidemiological Analysis of the Occurrence of Bovine Brucellosis and Adopted Control Measures in South Africa during the Period 2014 to 2019
Authors: Emily Simango, T. Chitura
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Background: Bovine brucellosis is among the most neglected zoonotic diseases in developing countries, where it is endemic and a growing challenge to public health. The development of cost-effective control measures for the disease can only be affirmed by the knowledge of the disease epidemiology and the ability to define its risk profiles. The aim of the study was to document the trend of bovine brucellosis and the control measures adopted following reported cases during the period 2014 to 2019 in South Africa. Methods: Data on confirmed cases of bovine brucellosis was retrieved from the website of the World Organisation of Animal Health (WOAH). Data was analysed using the Statistical Package for Social Sciences (IBM SPSS, 2022) version 29.0. Descriptive analysis (frequencies and percentages) and the Analysis of variance (ANOVA) were utilized for statistical significance (p<0.05). Results: The data retrieved in our study revealed an overall average bovine brucellosis prevalence of 8.48. There were statistically significant differences in bovine brucellosis prevalence across the provinces for the years 2016 and 2019 (p≥0.05), with the Eastern Cape Province having the highest prevalence in both instances. Documented control measures for the disease were limited to killing and disposal of disease cases as well as vaccination of susceptible animals. Conclusion: Bovine brucellosis is real in South Africa, with the risk profiles differing across the provinces. Information on brucellosis control measures in South Africa, as reported to the WOAH, is not comprehensive.Keywords: zoonotic, endemic, Eastern Cape province, vaccination
Procedia PDF Downloads 57217 Analysis of Microbiological Quality and Detection of Antibiotic Residue in Bovine Raw Milk Produced in Blida State, Algeria
Authors: M. N. Boukhatem, M. A. Ferhat, K. Mansour
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Bovine raw milk represents a favorable environment for the growth of several food-spoilage strains and some pathogens. It must meet stringent standards to ensure the highest microbiological and toxicological qualities.In order to assess the microbiological risks associated with the consumption of this food, we conducted this study to determine the microbiological quality of bovine raw milk (54 samples) commercialized at the state of Blida (Algeria). The samples analyzed were unsatisfactory in terms of total flora where 61.11% of samples were considered as non acceptable in terms of quality standards, fecal coliforms (40.74%), fecal streptococci (55.55%) and staphylococci (74.07%). Salmonella and Clostridium strains were not detected in all the samples. Furthermore, antibiotic residues were found in 26% of analysed samples. These results reflect non-compliance with the rules of good hygiene practices at milking, storage, transportatio, and sale of milk. Bovine raw milk consumed presents a serious health risk to the population of the study areas.The livestock coaching actors and dissemination of good hygiene practices throughout the production chain are needed to improve the quality of local milk.Keywords: bovine raw milk, microbiological quality, fecal coliforms, antibiotic residue, Blida state
Procedia PDF Downloads 228216 Obtaining High Purity Hydroxyapatite from Bovine Bone: Effect of Chemical and Thermal Treatments
Authors: Hernandez Pardo Diego F., Guiza Arguello Viviana R., Coy Echeverria Ana, Viejo Abrante Fernando
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The biological hydroxyapatite obtained from bovine bone arouses great interest in its application as a material for bone regeneration due to its better bioactive behavior in comparison with synthetic hydroxyapatite. For this reason, the objective of the present investigation was to determine the effect of chemical and thermal treatments in obtaining biological bovine hydroxyapatite of high purity and crystallinity. Two different chemical reagents were evaluated (NaOH and HCl) with the aim to remove the organic matrix of the bovine cortical bone. On the other hand, for analyzing the effect of thermal treatment temperature was ranged between 500 and 1000°C for a holding time of 4 hours. To accomplish the above, the materials before and after the chemical and thermal treatments were characterized by elemental compositional analysis (CHN), infrared spectroscopy by Fourier transform (FTIR), RAMAN spectroscopy, scanning electron microscopy (SEM), thermogravimetric analysis (TGA) and X-ray diffraction (XRD) and energy dispersion X-ray spectroscopy (EDS). The results allowed to establish that NaOH is more effective in the removal of the organic matrix of the bone when compared to HCl, whereas a thermal treatment at 700ºC for 4 hours was enough to obtain biological hydroxyapatite of high purity and crystallinity.Keywords: bovine bone, hydroxyapatite, biomaterials, thermal treatment
Procedia PDF Downloads 108215 Design of an Automatic Bovine Feeding Machine
Authors: Huseyin A. Yavasoglu, Yusuf Ziya Tengiz, Ali Göksenli
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In this study, an automatic feeding machine for different type and class of bovine animals is designed. Daily nutrition of a bovine consists of grass, corn, straw, silage, oat, wheat and different vitamins and minerals. The amount and mixture amount of each of the nutrition depends on different parameters of the bovine. These parameters are; age, sex, weight and maternity of the bovine, also outside temperature. The problem in a farm is to constitute the correct mixture and amount of nutrition for each animal. Faulty nutrition will cause an insufficient feeding of the animal concluding in an unhealthy bovine. To solve this problem, a new automatic feeding machine is designed. Travelling of the machine is performed by four tires, which is pulled by a tractor. The carrier consists of eight bins, which each of them carries a nutrition type. Capacity of each unit is 250 kg. At the bottom of each chamber is a sensor measuring the weight of the food inside. A funnel is at the bottom of each chamber by which open/close function is controlled by a valve. Each animal will carry a RFID tag including ID on its ear. A receiver on the feeding machine will read this ID and by given previous information by the operator (veterinarian), the system will detect the amount of each nutrition unit which will be given to the selected animal for feeding. In the system, each bin will open its exit gate by the help of the valve under the control of PLC (Programmable Logic Controller). The amount of each nutrition type will be controlled by measuring the open/close time. The exit canals of the bins are collected in a reservoir. To achieve a homogenous nitration, the collected feed will be mixed by a worm gear. Further the mixture will be transported by a help of a funnel to the feeding unit of the animal. The feeding process can be performed in 100 seconds. After feeding of the animal, the tractor pulls the travelling machine to the next animal. By the help of this system animals can be feeded by right amount and mixture of nutritionKeywords: bovine, feeding, nutrition, transportation, automatic
Procedia PDF Downloads 337214 Conservativeness of Functional Proteins in Bovine Milk by Pulsed Electric Field Technology
Authors: Sulhee Lee, Geon Kim, Young-Seo Park
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Unlike the traditional milk sterilization methods (LTLT, HTST, or UHT), pulsed electric field (PEF) technology is a non-thermal pasteurization process. This technology minimizes energy required for heat treatment in food processing, changes in sensory properties, and physical losses. In this study, structural changes of bovine milk proteins, the amount of immunoproteins such as IgG, and their storability by PEF treatment were examined. When the changes of protein content in PEF-treated milk were examined using HPLC, the amounts of α-casein and β-lactoglobulin were reduced over 40% each, whereas those of κ-casein and β-casein did not change. The amount of α-casein in HTST milk was reduced to 50%. When PEF was applied to milk at the energy level of 250 kJ, the amounts of IgG, IgA, β-lactoglobulin (β-LG), lactoferrin, and α-lactalbumin (α-LA) decreased by 43, 41, 35, 63, and 45%, respectively. When milk was sterilized by LTLT process followed by PEF process at the level of 150 kJ, the concentrations of IgG, IgA, β-LG, lactoferrin, and α-LA were 56.6, 10.6, 554, 2.8 and 660.1 μg/mL, respectively. When the bovine milk was sterilized by LTLT process followed by PEF process at the energy level of 180 kJ, storability of immunoproteins of milk was the highest and the concentrations of IgG, IgA, and β-LG decreased by 79.5, 6.5, and 134.5 μg/mL, respectively, when compared with the initial concentrations of those proteins. When bovine milk was stored at 4℃ after sterilization through HTST sterilizer followed by PEF process at the energy level of 200 kJ, the amount of lactoferrin decreased 7.3% after 36 days of storage, whereas that of lactoferrin of raw milk decreased 16.4%. Our results showed that PEF treatment did not change the protein structure nor induce protein denaturation in milk significantly when compared with LTLT or HTST sterilization. Also, LTLT or HTST process in combination with PEF were more effective than LTLT only or HTST only process in the conservation of immunoproteins in bovine milk.Keywords: pulsed electric field, bovine milk, immunoproteins, sterilization
Procedia PDF Downloads 431213 Staphylococcus argenteus: An Emerging Subclinical Bovine Mastitis Pathogen in Thailand
Authors: Natapol Pumipuntu
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Staphylococcus argenteus is the emerging species of S. aureus complex. It was generally misidentified as S. aureus by standard techniques and their features. S. argenteus is possibly emerging in both humans and animals, as well as increasing worldwide distribution. The objective of this study was to differentiate and identify S. argenteus from S. aureus, which has been collected and isolated from milk samples of subclinical bovine mastitis cases in Maha Sarakham province, Northeastern of Thailand. Twenty-one isolates of S. aureus, which confirmed by conventional methods and immune-agglutination method were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and multilocus sequence typing (MLST). The result from MALDI-TOF MS and MLST showed 6 from 42 isolates were confirmed as S. argenteus, and 36 isolates were S. aureus, respectively. This study indicated that the identification and classification method by using MALDI-TOF MS and MLST could accurately differentiate the emerging species, S. argenteus, from S. aureus complex which usually misdiagnosed. In addition, the identification of S. argenteus seems to be very limited despite the fact that it may be the important causative pathogen in bovine mastitis as well as pathogenic bacteria in food and milk. Therefore, it is very necessary for both bovine medicine and veterinary public health to emphasize and recognize this bacterial pathogen as the emerging disease of Staphylococcal bacteria and need further study about S. argenteus infection.Keywords: Staphylococcus argenteus, subclinical bovine mastitis, Staphylococcus aureus complex, mass spectrometry, MLST
Procedia PDF Downloads 141212 Influence of European Funds on the Sector of Bovine Milk and Meat in Romania in the Period 2007-2013
Authors: Andrei-Marius Sandu
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This study aims to analyze the bovine meat and milk sector for the period 2007-2013. For the period analyzed, it is known that Romania has benefited from EU funding through the National Rural Development Programme 2007-2013. In this programme, there were measures that addressed exclusively the animal husbandry sector in Romania. This paper presents data on bovine production of meat, milk and livestock in Romania, but also data on the price and impact the European Funds implementation had on them.Keywords: European funds, measures, national rural development programme, price
Procedia PDF Downloads 414211 Excision and Reconstruction of a Hypertrophic and Functional Bleb with Bovine Pericardium (Tutopatch®) and Amniotic Membrane: A Case Report
Authors: Blanca Fatela Cantillo, Silvia Iglesias Cerrato, Guadalupe Garrido Ceca
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Purpose: Bleb dysfunction is a late complication following glaucoma filtration surgery. We describe our surgical technique for excision and reconstruction of a hypertrophic bleb complication using bovine pericardium patch graft (Tutopatch®) and amniotic membrane. Material and methods: The case report presents a hypertrophic bleb over the cornea with good intraocular pressure control. The hanging bleb without leak caused dysesthesia and high irregular astigmatism. Bleb reconstruction involved the excision of corneal fibrous material and avascular conjunctiva, preserving the original scleral and tennon. Bovine pericardium patch graft (Tutopatch®) was sited over these with fixed sutures, reinforcing the underlying scleral, and the conjunctiva advanced. The superior epithelium corneal defect was covered using an amniotic membrane. Conclusion: Repair of bleb dysfunction with varied techniques has been reported, including conjunctival advancement, use of scleral patch graft, dural patch graft, or pericardium. Additional use of amniotic membrane promotes epithelialization and exhibits anti-fibrotic and anti-inflammatory features. Reconstruction with bovine pericardium patch graft and amniotic membrane resulted in pain relief, visual rehabilitation, and good aesthetic results, with preservation of bleb function.Keywords: reconstruction, hypertrophic bleb, bovine pericardium, amniotic membrane, dysesthesia of the bleb
Procedia PDF Downloads 69210 In vitro Effects of Berberine on the Vitality and Oxidative Profile of Bovine Spermatozoa
Authors: Eva Tvrdá, Hana Greifová, Peter Ivanič, Norbert Lukáč
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The aim of this study was to evaluate the dose- and time-dependent in vitro effects of berberine (BER), a natural alkaloid with numerous biological properties on bovine spermatozoa during three time periods (0 h, 2 h, 24 h). Bovine semen samples were diluted and cultivated in physiological saline solution containing 0.5% DMSO together with 200, 100, 50, 10, 5, and 1 μmol/L BER. Spermatozoa motility was assessed using the computer assisted semen analyzer. The viability of spermatozoa was assessed by the metabolic (MTT) assay, production of superoxide radicals was quantified using the nitroblue tetrazolium (NBT) test, and chemiluminescence was used to evaluate the generation of reactive oxygen species (ROS). Cell lysates were prepared and the extent of lipid peroxidation (LPO) was evaluated using the TBARS assay. The results of the movement activity showed a significant increase in the motility during long term cultivation in case of concentrations ranging between 1 and 10 μmol/L BER (P < 0.01; P < 0.001; 24 h). At the same time, supplementation of 1, 5 and 10 μmol/L BER led to a significant preservation of the cell viability (P < 0.001; 24 h). BER addition at a range of 1-50 μmol/L also provided a significantly higher protection against superoxide (P < 0.05) and ROS (P < 0.001; P < 0.01) overgeneration as well as LPO (P < 0.01; P<0.05) after a 24 h cultivation. We may suggest that supplementation of BER to bovine spermatozoa, particularly at concentrations ranging between 1 and 50 μmol/L, may offer protection to the motility, viability and oxidative status of the spermatozoa, particularly notable at 24 h.Keywords: berberine, bulls, motility, oxidative profile, spermatozoa, viability
Procedia PDF Downloads 120209 Intestinal Epithelium of Juvenile Dourado (Salminus brasiliensis) Fed with Bovine Colostrum
Authors: Thaline Maira P. Cruz, Debora B. Moretti, Wiolene M. Nordi, José Eurico P. Cyrino, Raul Machado-Neto
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The aim of this study was to evaluate the effect of lyophilized bovine colostrum (LBC) used as partial source of dietary protein on the histological characteristics of the intestinal epithelium of juvenile dourado (Salminus brasiliensis). Juveniles were fed with diets containing 0, 10 or 20% of lyophilized bovine colostrum (LBC) inclusion for either 30 or 60 days. For the histological study, the intestine was divided into three segments, S1, S2 and posterior intestine. In the S1 segment, interaction between treatment and period was observed in the number of goblet cells containing sialomucin, effect of treatment in the total number of goblet cells and effect of period in the number of goblet cells containing sulphomucins (P<0.05). In the S2 segment, effect of period was observed in the number of goblet cells containing acid, neutral and total mucins, sialomucins and Vv (P<0.05). In the posterior intestine, effect of period was observed in the thickness of muscle layer and number of goblet cells containing sialomucins and sulphomucins (P<0.05). Considering the aspects studied, the presence of lyophilized bovine colostrum in the diet did not significantly influence the enteric histological characteristics of juvenile dourado during the period of the study.Keywords: carnivorous fish, goblet cells, mucins, teleost
Procedia PDF Downloads 561