Alleviation Of Cyclic Heat Stress In Broilers By Dietary Supplementation Of Mannan-Oligosaccharides And Lactobacillus-based Probiotic
Material type: Book ; Format:
Publisher: 2011 Dissertation note: The antiviral activity of plants Silybum marianum (seeds), Chenopodium album (whole plant) and Nigella sativa (seeds) were evaluated against Peste des petitis ruminants virus (PPRV) and Foot and Mouth Disease virus (FMDV) in this study. Methanolic extraction of these plants was done by using Soxhlet apparatus and extracts were dried by using rotary evaporator. Six dilutions of each extracts 100, SO, 2S, 12.S, 6.2S, 3.12~g/ml were made in distilled water. Vero cells were infected by PPRV and BHK-21 by FMDV respectively. The herbal extracts assays of antiviral and cytotoxic were carried out in cell culture plates. Each well of 96 well cell culture plate were seeded with 104cell/ml of cell suspension. Cell counting was performed by hemocytometeric method. Positive and negative controls for antiviral and cytotoxic assay were also used, incubated the 96 well cell culture plates at 37°C for 4 days. After this incubation, MTT [3-(4,S-dimethylthiazol-2-yl)- 2,S-diphenyltetrazolium bromide] colorimetric assay were used for the determination of their quantification. Endpoint of this assay was considered in terms of cell survival percentage. Results were compared for qualitative variables using Chi-square technique and quantitative variables by linear regression analysis. 1 OO~g/ml and SOIlg/ml concentrations of Chenopodium album showed cell survival percentages of 87.9% and 86% respectively in PPRV and all six test dilutions of same plant showed no cytotoxicity for Vero cells. IOuug/ml and
SO~g/ml concentrations of Chenopodium album showed cell survival percentages of 88.5% and 87.2% respectively in FMDV and all six test dilutions of same plant showed no cytotoxicity for BHK-21 cells. Two concentrations of Nigella sativa 50!J. glml and 25!J. glml showed prominent cell s urvival of 85% and 84% respectively in PPRV and only one concentrations l Ouug/ml were found cytotoxic.Two concentrations of Nigella sativa 50uglml and 25!J.glml showed prominent cell survival of 79% and 77% respectively in FMDV and only one concentrations IOuug/ml were found cytotoxic. Only IOuug/ml of Silybum marianum has shown cytotoxicity and 50!J.glml and 25!J.glml shown prominent antiviral activity 91% and 85% respectively in PPRV. In FMDV l Otlug/ml of Silybum marianum has shown cytotoxicity and 50!J.glml and 25!J.g/ml shown prominent antiviral activity 93% and 91 % respectively. The results of present study are helpful in the treatment of Peste des petitis ruminants and Foot and Mouth Disease.
Availability: Items available for loan: UVAS Library [ Call number: 1360,T] (1).
Evaluation Of Different Extenders For The Cryopreservation Of Buffalo Bull (Bubalus Bubalis) Semen
Material type: Book ; Format:
; Literary form:
Publisher: 2011 Dissertation note: Buffalo is playing an important role in our country's economy by producing milk, meat and draught power. Genetic potential of low producing animals can be improved by using artificial insemination technology. Unfortunately, less number of elite bulls are available and low fertility rate of buffalo by using cryopreserved semen has been obtained. Semen is exposed to osmotic and oxidative stresses during processing, cryopreservation and thawing before insemination. Fertilizing ability is lost due to spermatozoa damage and it ultimately results in poor conception rates in buffalo. In order to protect spermatozoa from these stresses and improve fertility in buffalo, five osmotic pressure based concentrations of three extenders i.e. Citrate egg yolk extender (CEYE), Tris egg yolk extender (TEYE), and Lactose egg yolk extender (LEYE) were prepared by varying the quantity of the solutes to obtain an osmotic pressure of 255, 265, 275, 285 and 295 mOsm/kg. Osmotic pressure was measured by an osmometer.
In the first experiment, equal volume of semen obtained from four Nili-Ravi buffalo bulls was pooled and used to study the effects of osmotic pressure on post thawed semen characteristics. For this purpose, three basic media: citrate fructose media, tris citric acid fructose media and lactose media were prepared and divided each media in to five equal parts to maintain osmotic pressures of 255, 265, 275, 285 and 295mOsm/kg. These basis media were stored in a biomedical freezer, which were later used in preparing three semen extenders i.e. Citrate egg yolk extender (CEYE), Tris egg yolk extender (TEYE), and Lactose egg yolk extender (LEYE). During each collection, fifteen extenders (each of three extenders having five osmotic pressures i.e. 255, 265, 275, 285 and 295mOsm/kg) were used to extend the semen. After freezing, semen characteristics like sperm motility rate, viability rate, acrosomal integrity rate, plasma membrane integrity (PMI) rate, MTT reduction rate, sperm DNA integrity rate and lipid peroxidation were noted. Post thaw sperm motility rate in (%) CEYE was significant (P<0.05) at 295mOsm/kg compared to 255, 265 and 275mOsm/kg. However, sperm motility rate of different osmotic pressures of TEYE and LEYE was non-significant (P>0.05). Sperm viability rate (%) was non-significant (P>0.05) in all three extenders. Sperm acrosomal integrity rate was non-significant in CEYE and LEYE. However, it was significant (P<0.05) at 265, 275 and 295mOsm/kg in TEYE. Sperm PMI rate, MTT reduction rate, sperm DNA integrity rate and lipid peroxidation were non-significant (P>0.05) in CEYE, TEYE and LEYE.
On the basis of the individual and overall comparison of different semen characteristics of three extenders and their osmotic pressures, LEYE with 295mOs.kg was considered to be continued in the next experiment to upgrade the extender by adding taurine (TA) at 0.0, 30, 50 and 70 mM and trehalose (TR) at 0.0, 20, 40, 60 mM concentration. Semen collection, processing, freezing etc were done as per experiment-1 and same post thaw tests were carried out. Post thaw sperm motility rate was significantly (P<0.05) higher at TA-0.0 and TA-20mM and all concentration of TR. Sperm viability rate, acrosomal integrity rate, PMI rate, MTT reduction rate and lipid peroxidation at different concentrations of TA and TR were recorded non-significant (P>0.05). However, sperm DNA integrity rate was significant (P<0.05) higher at TA-0.0 and TR-0.0mM.
On the basis of comparison of different semen characteristics under various concentrations of taurine or trehalose, supplemented in semen extenders. Concentration of TR-70mM was considered to be continued in the next experiment to test fertility of the optimized extender.
Semen straws of LEYE supplemented with TR-70mM were used to inseminate the 50 buffaloes in heat (Supplemented group), while, traditionally used tris based buffalo bull semen extender was used (control group) to compared pregnancy rate (PR) of this experiment. Pregnancy rate in control and supplemented group was 38 and 54% respectively, which was statistically non-significant (P>0.05).
Availability: Items available for loan: UVAS Library [ Call number: 1538,T] (1).
Seasonal Variations In Libido Semen Quality Freezability And Fertility Of Cholistani Bulls
Material type: Book ; Format:
; Literary form:
Publisher: 2013 Dissertation note: Abstract
Availability: Items available for loan: UVAS Library [ Call number: 1901,T] (1).
Response Of Dietary Supplementation Of Β-Galacto-Oligosaccharides And Benzoic Acid On Growth Performance And Gastrointestinal Physiology In Broiler Chickens
Material type: Book ; Literary form:
Publisher: 2015 Dissertation note: Efficient poultry production depends upon a number of crucial factors that include optimum nutrient digestibility, bioavailability and maximum conversion into end products like meat and egg as well as low morbidity and mortality in the flock. In the past, antibiotics were used successfully as growth promoters in poultry and animal feed to improve production performance. However, it has become increasingly apparent that many problems are associated with the use of antibiotic growth promoters (AGPs); most importantly, the emergence of increasing numbers of microbes resistant to antibiotics. Therefore, AGPs were banned in the poultry as well as in the livestock industries of many countries. Due to withdrawal of AGPs, the poultry industry faced outbreak of diseases, high mortality, compromised growth performance and thus low profitability. Researchers and producers must, therefore, seek an alternate for AGPs that provide safety both for poultry and consumer health. Many potential candidates have subsequently been put forth and evaluated to replace AGPs that include prebiotics, probiotics, phytobiotics and organic acids. These AGPs replacers showed variable degree of success depending upon nutritional, managemental and hygienic conditions. Keeping in view the importance of AGP replacers, it was the purpose of this study to the growth promoting potential of β-galacto-oligosaccharides (β-GOS), a prebiotic, and benzoic acid (BA), an organic acid, in growing broilers.
In experiment-1, 200 day old Hubbard broiler chicks, were randomly divided into four treatment groups (n = 50) with five replicates (n = 10) in each group. Birds in different treatment groups were allotted four different types of diets. Birds in the control group (β-GOS-0) were given ad-libitum corn based basal diet, free of antimicrobials and coccidiostats, whereas birds in three other experimental groups were given the same basal diets supplemented with different concentrations of β-GOS i.e., 0.1% (β-GOS-1), 0.2% (β-GOS-2) and 0.5% (β-GOS-5) for a period of 35 days. Birds in all the experimental groups were vaccinated against the Newcastle disease virus (NDV) and infectious bursal disease virus by giving primary and booster doses according to the schedule. Moreover, birds were also sensitized against 2, 4-dinitrochlorobenzene for determination of cell mediated immunity (CMI). Body weights, feed intake and feed conversion efficiency were determined on a weekly basis. At the end of experiment, ten birds from each group were slaughtered to collect blood, organs and caecal digesta for determination of serum biochemical metabolites and humoral immunity, organ characteristics and selected caecal digesta bacteria, respectively. Data were analyzed by using analysis of variance and incase of significant F-value (P < 0.05), data were subjected to Tukey's-HSD post hoc test. Results demonstrated that growth performance and feed conversion efficiency (FCE) of β-GOS supplemented birds were improved (P < 0.05) in a dose dependent manner, with the highest body weights (P < 0.05) and improved FCE (P < 0.05) observed in the β-GOS-5 group. Relative weights of liver and pancreas were higher (P < 0.05) in the β-GOS-1 group. Likewise, CMI was also higher (P < 0.05) in the β-GOS-1 group. Concentrations of caecal Lactobacilli spp. were higher in the β-GOS-2 and β-GOS-5 groups. The serum biochemical metabolites, humoral immunity in terms of antibodies titres against NDV and Clostridia spp. and coliforms were not affected by dietary supplementation of β-GOS. In conclusion, dietary supplementation of 0.5% β-GOS conferred a better response in terms of growth performance, feed conversion efficiency and caecal Lactobacilli spp. in broilers.
In experiment-2, 120 day old male Cobb broiler chicks were randomly divided into three treatment groups (n = 40) with eight replicates (n = 5) in each group. Birds in different treatment groups were allotted three different types of diets. Birds in the control group (CON) were given ad-libitum maize based basal diets, free of antimicrobials and coccidiostats, whereas birds in two other experimental groups were given the same basal diets supplemented with two different types of BA i.e., 0.096% free BA (FBA) and 0.2% slow releasing BA (SBA) for a period of 35 days. Body weights, feed intake and feed conversion efficiency were determined on a weekly basis. At the end of the experimental period, birds from each group were slaughtered to collect digesta from the crop, proventriculus, gizzard, proximal small intestine, ileum and caecum for determination of benzoic acid, microbial metabolites, microbial populations and molecular microbial ecology. Data were analyzed by using analysis of variance and incase of significant F-value (P < 0.05), data were subjected to Tukey's-HSD post hoc test. Results revealed that overall growth performance, feed intake and FCE remained unchanged among all the treatment groups. Total short chain fatty acids were only increased (P < 0.05) in the gizzard by both types of BA supplementation. Likewise, D-lactate concentration was increased (P < 0.05) by both types of BA in crop digesta, whereas, L-lactate was only increased (P < 0.05) by SBA in crop digesta. Both types of BA increased (P < 0.05) the concentrations of L-lactate, D-lactate and acetic acid in gizzard, and D-lactate in proximal small intestine. Concentrations of i-butyric acid and i-valeric acid were decreased (P < 0.05) in caecal digesta by both types of BA, whereas, n-valeric acid was only decreased (P < 0.05) by SBA supplementation in caecal digesta. Concentrations of both types of BA in different segments of gastrointestinal tract were comparable. Concentrations of gizzard Lactobacilli spp. were higher in both the BA supplemented groups, whereas, L. johnsoni and L. reuteri concentrations were higher in the SBA group compared with the control group. Similarly, in the proximal small intestine both types of BA increased the population of Lactobacilli spp., L. reuteri, L. amylovorus and Enterococci spp. Ileal concentrations of Lactobacilli spp., L. johnsoni, L. amylovorus and S. alactolyticus were higher (P < 0.05) in SBA group, whereas, L. reuteri was increased (P < 0.05) both by the FBA and SBA supplementations. Microbial populations of crop and caecum remained unchanged among all the treatment groups. Results of ileal molecular microbial ecology analysis revealed that evenness was higher (P < 0.05) in both the BA supplemented groups, whereas, the Sorensen index (P < 0.05) and the Renkonen index (< 0.5) were different only in the SBA group. However, caecal microbial ecology analysis revealed no changes among all the treatment groups. In conclusion, FBA and SBA supplementations produced similar responses in terms of microbial metabolites production with no effect on overall growth performance and feed conversion efficiency. However, SBA conferred some advantages over FBA in terms of increased bacterial concentrations, especially some species of the Lactobacilli as well as S. alactolyticus, and improved ileal molecular microbial ecology.
Availability: Items available for loan: UVAS Library [ Call number: 2310-T] (1).
Development Of A Suitable Semen Extender For The Cryopreservation Of Nili Ravi Buffalo Bull (Bubalus Bubalis) Semen
Material type: Book ; Literary form:
Publisher: 2015 Dissertation note: Presently, buffalo farmers are dissatisfied with fertility rates of the frozen semen used in the field and tend to use bulls. This study was designed to develop a suitable semen extender for cryopreservation of Nili Ravi buffalo semen that can improve conception rate in buffaloes.
Experiment-I, an attempt was made to develop semen extender with optimal osmotic pressure for buffalo semen using tris citric acid (TCAE), skim milk (SME) and coconut water (CWE) extenders (each extender have 260, 270, 280, 290 and 300 mOsm/kg osmotic pressure levels). In Experiment-II, best extender (TCAE: 300 mOsm/kg) of experiment-I was tried to improve post thaw spermatozoa characteristics by supplementing antioxidants [0.0, 1.75, 2.0 and 2.25 mM butylated hydroxy toluene (BHT) and 0.0, 2.0, 5.0 and 8.0 mM L-cysteine]. Post thaw spermatozoa motility, viability, plasma membrane integrity (PMI), DNA damage rate and lipid peroxidation were assessed in first two experiments. In Experiment-III, pregnancy rate assessment of extended semen was carried out by using Trial extender (best of experiment II) or Control extender of Semen Production Unit (SPU), Qadirabad, Pakistan (50 inseminations of each extender).
Higher spermatozoa motility at ≥ 270 mOsm/kg was noted in TCAE than both SME and CWE could be due to less intracellular ice formation in zwitterions extender. Higher spermatozoa viability in TCAE and CWE compared to SME may be attributed to extender effectiveness. Higher acrosomal integrity rate at 300 mOsm/kg in TCAE and SME may be because of less intracellular ice formation in isotonic extenders. At 290 mOsm/kg, higher spermatozoa PMI in SME and lesser DNA damage in three extenders might be due to lesser intracellular ice formation at cryopreservation. Decreased spermatozoa DNA damage in SME might be due to the presence of natural antioxidants i.e., casein. Higher lipid peroxidation in CWE than TCAE and SME may be due to presence of natural antioxidants (in SME) and higher cell dehydration potential of TCAE.
Higher spermatozoa motility recorded at 2.0 mM BHT compared to other BHT groups including DMSO might be due to fact that BHT protects spermatozoa mitochondria by reducing oxidative stress. Lower spermatozoa viability, PMI rates and higher DNA damage at 2.25 mM of BHT may be due to BHT toxic effects. Lower lipid peroxidation in BHT treated groups compared to DMSO and BHT control groups might be related to BHT strong antioxidant properties. L-cysteine caused higher spermatozoa DNA damage at highest level (i.e., 8 mM) that could also be due to antioxidant’s toxic effect.
Pregnancy rate 18 % higher was noted in Trial than Control semen extender; however no significant difference have been noted that might be due to less no of inseminations.
In conclusion, TCA extender (300 mOsm/kg) having BHT (2.0 mM) improved post thaw semen quality and yielded numerically better pregnancy rates. Results of study indicated that osmotic stress damaged the spermatozoa internal structures more severely than injury to plasma membrane.
Availability: Items available for loan: UVAS Library [ Call number: 2360-T] (1).