Species adulteration in raw milk samples using polymerase chain reaction-restriction fragment length polymorphism

Research (Published online: 22-06-2018)

15. Species adulteration in raw milk samples using polymerase chain reaction-restriction fragment length polymorphism

Rania M. Ewida and Doaa Safwat M. Abd El-Magiud

Veterinary World, 11(6): 830-833

ABSTRACT

Background and Aim: Milk adulteration is pivotal because it leads to worse effects in public health as human adverse reactions with clinical signs ranged from gastrointestinal signs to anaphylactic shock. This study was carried out to estimate the prevalence of adulteration in buffalo's milk sold in Assiut City, Egypt.

Materials and Methods: A total of 50 raw buffalo's milk samples were collected and examined for adulteration by addition of cow's milk. The examination carried out by applying polymerase chain reaction-restriction fragment length polymorphism technique using cytochrome b (cyt b) gene primers and Hinf I enzymes. The size of target gene was 360 bp in both animal species and amplicon can be digested using Hinf I enzyme, this restriction enzyme divided the essential band to clear three bands at 360, 210, and 150 bp in cows' milk, while, the enzyme could not be cleaved the amplicon in buffalo's samples.

Results: The obtained results cleared that the incidence of adulteration of buffalo's milk very high percentage reaches 90%.

Conclusion: It could be concluded that the raw buffalo's milk sold in Assiut City subject to fraudulent practice and thus can lead to public health hazards.

Keywords: buffalo's milk, cyt b gene, Hinf I enzyme, medicolegal, milk adulteration, polymerase chain reaction-restriction fragment length polymorphism.

Antiviral activity of Acanthaster planci phospholipase A2 against human immunodeficiency virus

Research (Published online: 20-06-2018)

14. Antiviral activity of Acanthaster planci phospholipase A2 against human immunodeficiency virus

Anondho Wijanarko, Kenny Lischer, Heri Hermansyah, Diah Kartika Pratami and Muhamad Sahlan

Veterinary World, 11(6): 824-829

ABSTRACT

Aim: Investigation of antiviral activity of Acanthaster planci phospholipase A2 (AP-PLA2) from moluccas to human immunodeficiency virus (HIV).

Materials and Methods: Crude venom (CV) and F20 (PLA2 with 20% fractioned by ammonium sulfate) as a sample of PLA 2 obtained from A. planci's extract were used. Enzymatic activity of PLA2 was determined using the degradation of phosphatidylcholine (PC). Activity test was performed using in vitro method using coculture of phytohemagglutinin-stimulated peripheral blood mononuclear cell (PBMC) from a blood donor and PBMC from HIV patient. Toxicity test of AP-PLA2 was done using lethal concentration required to kill 50% of the population (LC50).

Results: AP-PLA2 F20 had activity and purity by 15.66 times bigger than CV. The test showed that the LC50 of AP-PLA2 is 1.638 mg/ml. Antiviral analysis of AP-PLA2 in vitro showed the inhibition of HIV infection to PBMC. HIV culture with AP-PLA2 and without AP-PLA2 has shown the number of infected PBMC (0.299±0.212% and 9.718±0.802%). Subsequently, RNA amplification of HIV using reverse transcriptase-polymerase chain reaction resulted in the decrease of band intensity in gag gene of HIV.

Conclusion: This research suggests that AP-PLA2 has the potential to develop as an antiviral agent because in vitro experiment showed its ability to decrease HIV infection in PBMC and the number of HIV ribonucleic acid in culture.

Keywords: Acanthaster planci, antiviral activity, human immunodeficiency virus, Indonesia, phospholipase.

Humoral and cellular immune response induced by antigenic protein of Sarcoptes scabiei var. caprae

Research (Published online: 19-06-2018)

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

ABSTRACT

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.

Keywords: antigenic protein, CD4, CD8, immunoglobulin G, Sarcoptes scabiei var. caprae, toll-like receptor-9.

Methane mitigation potential of phyto-sources from Northeast India and their effect on rumen fermentation characteristics and protozoa in vitro

Research (Published online: 19-06-2018)

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.

Keywords: methane, phyto-sources, protozoa, rumen fermentation, tannin.

The effectiveness of platelet-rich plasma on the skin wound healing process: A comparative experimental study in sheep

Research (Published online: 14-06-2018)

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

ABSTRACT

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.

Keywords: healing, platelet-rich plasma, sheep, skin

Protective role of Brucella abortus specific murine antibodies in inhibiting systemic proliferation of virulent strain 544 in mice and guinea pig

Research (Published online: 13-06-2018)

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

ABSTRACT

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.

Keywords: brucellosis, humoral immunity, mice, passive protection, protective index.

Threshold somatic cell count for delineation of subclinical mastitis cases

Research (Published online: 12-06-2018)

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

ABSTRACT

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.

Keywords: discriminate function, mastitis, receiver's operating characteristic curve, somatic cell count, threshold.