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13. Humoral and cellular immune response induced by antigenic protein of Sarcoptes scabiei var. caprae
Nunuk Dyah Retno Lastuti, Wiwik Misaco Yuniarti, Poedji Hastutiek, Lucia Tri Suwanti and Dony Chrismanto
Veterinary World, 11(6): 819-823
Aim: Scabies is one of the most important diseases in goats and caused by a complex hypersensitivity process that involves both humoral and cell-mediated immune responses. This phenomenon shows that the variety of Sarcoptes scabiei has different characteristics of specific antigenic protein or different immune-dominant. This research aims to detect the humoral and cellular immune response of rabbits which were immunized with the protein of S. scabiei var. caprae.
Materials and Methods: This research was done as follows, identification and collection of Sarcoptes scabiei var caprae from goat infected with scabies, separation of protein antigen from S. scabiei mites with ultrasonic sonicator, measurement of protein content with spectrophotometry, rabbit injection with 500 μg dose of antigen protein which was repeated 5 times (5x booster) every 2 weeks. Measurement of IgG titer using indirect ELISA, whereas to detect the expression of cellular immune response (TLR-9, CD4, and CD8) using Direct Immunofluorescence assay.
Results: Based on the statistical analysis, it showed that there was a significant enhancement (p<0.05) for optical density value or antibody titer and cellular immune response was shown by TLR-9, CD4, and CD8 expression in rabbit T lymphocytes which appear yellow to green fluorescent color using fluorescence microscope. The amount of fluorescence T lymphocytes showed a significant difference (p<0.05) between the control and various boosters.
Conclusion: Antigenic protein of S. scabiei var. caprae contains ligands, which are involved in the pathogen-associated molecular pattern that has an ability to induce humoral and cellular immune response in rabbit. Specifically, that TLR-9 is not only involved in innate immunity but also in adaptive immunity and can be used as alternative adjuvant development research.
12. Methane mitigation potential of phyto-sources from Northeast India and their effect on rumen fermentation characteristics and protozoa in vitro
Luna Baruah, Pradeep Kumar Malik, Atul P. Kolte, Arindam Dhali and Raghavendra Bhatta
Veterinary World, 11(6): 809-818
Aim: The aim of the study was to explore the anti-methanogenic potential of phyto-sources from Northeast region of the country and assess the effect on rumen fermentation characteristics and protozoa for their likely inclusion in animal diet to reduce methane emission.
Materials and Methods: Twenty phyto-sources were collected from Northeast state, Assam, during March to April 2014. Phyto-sources were analyzed for their tannin content followed by screening for methane mitigation potential using in vitro system. The effect of tannin on methane production and other fermentation parameters was confirmed by attenuating the effect of tannin with polyethylene glycol (PEG)-6000 addition. About 200 mg dried phyto-source samples were incubated for 24 h in vitro, and volume of gas produced was recorded. The gas sample was analyzed on gas chromatograph for the proportion of methane in the sample. The effect of phyto-sources on rumen fermentation characteristics and protozoal population was determined using standard methodologies.
Results: Results from studies demonstrated that Litchi chinensis, Melastoma malabathricum, Lagerstroemia speciosa, Terminalia chebula, and Syzygium cumini produced comparatively less methane, while Christella parasitica, Leucas linifolia, Citrus grandis, and Aquilaria malaccensis produced relatively more methane during in vitro incubation. An increase (p<0.05) in gas and methane production from the phyto-sources was observed when incubated with PEG-6000. Entodinimorphs were prominent ciliates irrespective of the phyto-sources, while holotrichs represented only small fraction of protozoa. An increase (p<0.05) in total protozoa, entodinimorphs, and holotrichs was noted when PEG-6000 added to the basal substrate. Our study confirmed variable impact of phyto-sources on total volatile fatty acid production and ammonia-N.
Conclusion: It may be concluded that L. chinensis, M. malabathricum, L. speciosa, S. cumini, and T. chebula are having potent methane suppressing properties as observed in vitro in 24 h. These leaves could be supplemented in the animal diet for reducing methane emission; however, in vivo trials are warranted to confirm the methane inhibitory action and optimize the level of supplementation.
11. The effectiveness of platelet-rich plasma on the skin wound healing process: A comparative experimental study in sheep
Daikh Badis and Bennoune Omar
Veterinary World, 11(6): 800-808
Aim: The therapeutic evaluation of the biological effect of platelet-rich plasma (PRP) used as a surgical adjunct to maintain the inflammatory process and to potentiate tissue healing, make the subject of recent research in regenerative medicine. This study was designed to evaluate the healing activity of PRP by its topical application on the skin experimentally injured in a sheep model.
Materials and Methods: The study was conducted on 9 adult and clinically healthy males sheep. PRP was obtained by a protocol of double centrifugation of whole blood from each animal. After sterile skin preparation, full-thickness excisional wounds (20 mm x 20 mm) were created on the back of each animal. The animals were randomly divided into three equal groups of three sheep for each. In Group I, the wounds were treated with PRP, in Group II; wounds were treated with Asiaticoside; in Group III, wounds were treated with saline solution. The different treatments were administered topically every 3 days. Morphometric measurements of the contraction surface of the wounds and histopathological biopsies were carried out at the 3rd, 7th, 14th, 21st, and 28th days of healing.
Results: The results of the morphometric data obtained revealed that it was significant differences recorded at the 7th and 14th day of healing in favor for animals of Group I. Semi-quantitative histopathological evaluation showed that PRP reduces inflammation during 3 first days post-surgical and promotes epithelialization in 3 weeks of healing.
Conclusion: We concluded that topical administration of PRP obtained by double centrifugation protocol could potentially improve the skin healing process in sheep.
10. Protective role of Brucella abortus specific murine antibodies in inhibiting systemic proliferation of virulent strain 544 in mice and guinea pig
Suman Verma, Mayank Rawat, Sanjay Kumawat, Salauddin Qureshi, Gulam Mohd and Ashok Kumar Tiwari
Veterinary World, 11(6): 794-799
Aim: The major objective of the investigation was to evaluate the hitherto uncharacterized potential of Brucella-specific antibodies to win the battle against virulent Brucella abortus infection.
Materials and Methods:Brucella-specific immune serum was raised in mice. The antibody titer of serum was determined by standard tube agglutination test and indirect enzyme-linked immunosorbent assays (iELISA). Groups of mice and guinea pigs were passively immunized with serum containing specific agglutinin titers. 24 h after immunization, all animals along with unimmunized controls were challenged with B. abortus S544. Total B. abortus S544 counts in the spleen of each animal collected on the 7th day of challenge was determined to evaluate the protective index (PI) of anti-Brucella serum by statistical analysis.
Results: A dose-dependent protective response to immune mice serum was observed in both experimental models though the values of PI of mice were higher than those obtained for guinea pigs. The PI values in mice passively immunized with 50 IU or 25 IU antibodies were 1.38 and 0.69, respectively. In guinea pigs, however, animals passively immunized with 50 IU or 25 IU antibodies showed PI values equivalent to 0.79 and 0.41, respectively.
Conclusion: The observations support our hypothesis that the presence of antibodies inhibits the initial multiplication and eventual colonization of systemic organs by B. abortus. Therefore, a predominant antibody-mediated response induced by a vaccine is expected to protect the animal against the most severe clinical outcome of infection.
9. Threshold somatic cell count for delineation of subclinical mastitis cases
P. V. Jadhav, D. N. Das, K. P. Suresh and B. R. Shome
Veterinary World, 11(6): 789-793
Aim: Somatic cell count (SCC) is the most widely used single reliable indicator of udder health. The present study was carried out with an objective to find the exact threshold of SCC.
Materials and Methods: Milk samples collected from a total of 214 Holstein Friesian crossbred dairy animals were subjected to bacterial DNA extraction and SCC estimation by digital PortaCheck. California Mastitis Test and polymerase chain reaction based on amplification of organism using reported primers were performed to diagnose subclinical mastitis. Receiver's operating characteristic (ROC) curve analysis and discriminate function analyses were performed using SPSS 18 software.
Results: ROC curve analysis represented that the area under the curve was 0.930 with the standard error of 0.02. Results indicated that 93% of the case could be correctly predicted as mastitis infected using SCC as a marker (p<0.001). At cut score level of 282 000 cells/ml, 285,000 cells/ml and 288,000 cells/ml, sensitivity remained 92.6% and specificity augmented as 86.3%, 87.2%, and 88%, respectively. At SCC value of 310,000 cells/ml of milk, sensitivity and specificity were optimal, namely, 92.6% and 91.5%, respectively. The function fitted demonstrated 89.2% accuracy with p<0.001. The functions at group centroids were -0.982 and 1.209, respectively, for normal and mastitis-infected animals and log_SCC value was the most important factor contributing 38.30% of the total distance measured.
Conclusion: Our study supports that the threshold value to delineate subclinical mastitis case from the normal is 310,000 somatic cells/ml of milk and a model so fitted using the variable SCC can be successfully used in field for the diagnosis of subclinical cases of mastitis which otherwise would be difficult to differentiate based on clinical signs.
8. Astaxanthin inhibits cytokines production and inflammatory gene expression by suppressing IκB kinase-dependent nuclear factor κB activation in pre and postpartum Murrah buffaloes during different seasons
Lakshmi Priyadarshini and Anjali Aggarwal
Veterinary World, 11(6): 782-788
Aim: We examined regulatory function of astaxanthin on mRNA expression of nuclear factor κB (NF-κB) p65, interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-α), and interferon gamma (IFN-γ) in peripheral blood mononuclear cells in pre and postpartum Murrah buffaloes during summer (temperature-humidity index [THI]=86; relative humidity [RH]=24) and winter (THI=58.74; RH=73) seasons.
Materials and Methods: A total of 32 Murrah buffaloes apparently healthy and in their one to four parity were selected from National Dairy Research Institute herd and equally distributed randomly into four groups (control and supplemented groups of buffaloes during summer and winter season, respectively). All groups were fed according to the nutrient requirement of buffaloes (ICAR, 2013). The treatment group was supplemented with astaxanthin at 0.25 mg/kg body weight/animal/day during the period 30 days before expected date of calving and up to 30 days postpartum.
Results: There was downregulation of NF-κB p65 gene in all the groups. NF-κB p65 mRNA expression was lower (p<0.05) in treatment than control group from prepartum to postpartum during summer, while mRNA expression was low only on day 21 after calving during winter season. The mRNA expression of IL-6, TNF-α, and IFN-γ was lower (p<0.05) in treatment than a control group of buffaloes during summer and winter seasons. The mRNA expression of NF-κB p65, IL-6, TNF-α, and IFN-γ was higher (p<0.05) in summer than in winter seasons.
Conclusion: The xanthophyll carotenoid astaxanthin a reddish-colored C-40 compound is a powerful broad-ranging antioxidant that naturally occurs in a wide variety of living organisms, such as microalgae, fungi, crustaceans, and complex plants. Astaxanthin blocked nuclear translocation of NF-κB p65 subunit and IκBa degradation, which correlated with its inhibitory effect on IκB kinase (IKK) activity. These results suggest that astaxanthin, probably due to its antioxidant activity, inhibits the production of inflammatory mediators by blocking NF-κB activation and as a consequent suppression of IKK activity and IκB-degradation.
7. Genetic diversity and population structure of local and exotic sheep breeds in Jordan using microsatellites markers
Khaleel I. Jawasreh, Mustafa M. Ababneh, Zuhair Bani Ismail, Abdel Mon'em Bani Younes and Ibrahem Al Sukhni
Veterinary World, 11(6): 778-781
Aim: This study was conducted to study the genetic and population structure of local (Awassi) and exotic (Romanov, Charollais, Assaf, Awassi, and Suffolk) sheep breeds in Jordan using eight microsatellite markers.
Materials and Methods: A total of 125 sheep were used (25 from each breed) in the study. The number of alleles (A), the mean values of observed (Ho) and expected (He) heterozygosity, polymorphism information content (PIC), fixation index as a measure of heterozygote deficiency or excess, and Hardy-Weinberg equilibrium (HWE) were analyzed using PopGen and CERVUS softwares. Nei's standard genetic distances among breeds and dendrogram of unweighted pair group method with arithmetic mean (UPGMA) were calculated and constructed using PopGen software.
Results: A total of 40 alleles were detected with an average number of alleles of 5. The mean Ho value was higher than the mean He value for all breeds. Awassi breed showed the highest average PIC value while Romanov had the lowest. There was a significant (p<0.05) deviation from HWE at each locus within and between breeds. Deviations from HWE were found to be highly significant for all markers except OARFCP304 locus. The genetic distance estimates revealed a close relationship between Romanov and Charollais and between Awassi and Charollais. In the UPGMA dendrogram, Charollais, Romanov, and Awassi breeds were placed together in one main cluster while Assaf was in a different subcluster. Awassi was placed alone in a second main cluster.
Conclusion: Results of this study offer insight toward the genetic conservation of the studied breeds and a base on which breeding plans can be made.