Detection, identification, and differentiation of sheep pox virus and goat pox virus from clinical cases in Giza Governorate, Egypt

Research (Published online: 18-12-2016)

18. Detection, identification, and differentiation of sheep pox virus and goat pox virus from clinical cases in Giza Governorate, Egypt - M. A. Mahmoud and M. H. Khafagi

Veterinary World, 9(12): 1445-1449

   doi: 10.14202/vetworld.2016.1445-1449

Abstract

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Aim: To isolate, identify, and differentiate Capripoxviruses (CaPV) (sheep pox virus and goat pox virus) infections by egg inoculation, transmission electron microscopy (TEM), and 30 kDa RNA polymerase subunit gene-based polymerase chain reaction (PCR) (RPO30) in clinically affected animals in Hawamdia township of Giza Governorate, Egypt.

Materials and Methods: A total of 37 scab samples were collected from clinically suspected field cases of sheep pox and goat pox. These samples were collected during (2014-2015) during different outbreaks of sheep pox and goat pox from Hawamdia township of Giza Governorate, Egypt. The samples were subjected to egg inoculation, TEM, and (RPO30) gene-based PCR. By using the egg inoculation: Previously prepared 37 scab samples (n=23 sheep and n=14 goats) were inoculated on the chorioallantoic membrane of specific pathogen free (SPF) embryonated chicken eggs (12 days old age). In the presence of the suitable percentage of humidity and candling, the inoculated eggs were incubated at 37°C. By using the TEM: Samples showed positive pock lesions on the chorioallantoic membranes, were fixed in glutaraldehyde, then processed and sectioned for TEM. Using the (RPO30) gene-based PCR assay, 30 of positive samples after egg inoculation (n=19 sheep and n=11 goats) were screened.

Results: Using the egg inoculation, a characteristic pock lesions for poxviruses were seen in 30/37 (n=19 sheep and n=11 goats) (81.08%). Using the TEM, examination of the positive samples after egg inoculation revealed positive result in 23/30 (n=15 sheep and n=8 goats) (76.66%). The positive results represented by the presence of negatively stained oval-shape virus particles. Using the (RPO30) gene-based PCR assay, out of 30 total of positive samples after egg inoculation (n=19 sheep and n=11 goats) were screened, 27 (90%) samples (n=17 sheep and n=10 goats) were positive. The given band sizes of sheep and goats were 172 and 152 bp, respectively.

Conclusion: PCR assay depended on RPO30 gene can be used lonely for the detection, identification, and differentiation of CaPVs. RPO30 gene-based PCR assay in combination with gene sequencing helps in molecular epidemiological studies of CaPV infection.

Keywords: Capripoxvirus, polymerase chain reaction, goat pox, isolation, RPO30 gene, sheep pox, transmission electron microscope.

Evaluation of sperm recovered after slaughter from cauda epididymides of red Sokoto bucks

Research (Published online: 18-12-2016)

17. Evaluation of sperm recovered after slaughter from cauda epididymides of red Sokoto bucks - A. H. Abu, A. I. Kisani and T. Ahemen

Veterinary World, 9(12): 1440-1444

   doi: 10.14202/vetworld.2016.1440-1444

Abstract

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Aim: Viable spermatozoa could be recovered from the cauda epididymides for the purpose of preservation of genetic material of male animals with desirable traits and for use in reproductive biotechnology. The aim of this study was to determine the effect of storage time on testicular and epididymal biometry, sperm reserves and epididymal sperm characteristics of red Sokoto bucks post mortem.

Materials and Methods: Testes-epididymides were collected immediately after slaughter of mature red Sokoto bucks and transported in ice chest to the laboratory. The samples were either processed immediately or stored at 4°C in refrigerator for 24, 48 h and then processed. The testes and epididymides were measured and weighed. Sperm motility, concentration, livability, morphology, intact acrosome, and sperm reserves from different treatment groups including control were evaluated and means (±standard error of mean) were recorded.

Results: There was no significant difference (p>0.05) in the testicular and epididymal dimensions determined between the means of the groups. Percent sperm motility and viability decreased significantly (p<0.05) after 24 h from 69.00±0.46 and 71.27±0.50% to 50.60±0.48 and 60.47±0.70% at 48 h, respectively. Significant decreases (p<0.05) in epididymal sperm concentration and intact acrosome from 2.86±0.08 and 92.87±0.39 at 0 to 24 h of storage, respectively, were observed. Gonadal sperm reserves and sperm reserves in the cauda epididymides decreased significantly (p<0.05) from 0 to 48 h of cold storage. However, no significant decrease (p>0.05) in caput and corpus sperm reserves was observed from 0 to 48 h of storage.

Conclusion: The results of this study suggest that spermatozoa recovered from the epididymides of red Sokoto bucks were viable after storage for up to 48 h. Furthermore, this finding offers some hope that epididymal sperm recovered post-mortem can be used in assisted reproductive technologies.

Keywords: epididymides, goat bucks, spermatozoa, testes.

Phenotypic and genotypic characterization of locally isolated Salmonella strains used in preparation of Salmonella antigens in Egypt

Research (Published online: 17-12-2016)

16. Phenotypic and genotypic characterization of locally isolated Salmonella strains used in preparation of Salmonella antigens in Egypt -Hazem Mohammed Ibrahim, Dalia Mohammed Adb-Elmoaty, Hanan Ali Ahmed and Mona Ibrahim El-Enbaawy

Veterinary World, 9(12): 1435-1439

   doi: 10.14202/vetworld.2016.1435-1439

Abstract

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Aim: This work was conducted to study the phenotypic and genotypic characterization of locally isolated Salmonella strains (SalmonellaPullorum, Salmonella Enteritidis, and Salmonella Typhimurium) from poultry used in the preparation of Salmonella antigens in Egypt.

Materials and Methods: The phenotypic characterization of Salmonella strains was done using standard microbiological, biochemical, and serological techniques. Molecular identification was done using different sets of primers on different genes using different polymerase chain reaction (PCR) techniques.

Results: The phenotypic characterization of Salmonella strains was confirmed. Molecular identification revealed detection of 284 bp fragment of InvA gene in all studied Salmonella strains. Furthermore, multiplex PCR was used for more confirmation of being Salmonella spp., generally at 429 bp as well as genotyping of Salmonella Typhimurium and Salmonella Enteritidis at 559 and 312 bp, respectively, in one reaction.

Conclusion: The locally isolated field Salmonella strains were confirmed phenotypically and genotypically to be Salmonella Enteritidis, and Salmonella Typhimurium and could be used for the preparation of Salmonella antigens.

Keywords: characterization, duplex polymerase chain reaction, multiplex polymerase chain reaction, Salmonella spp.

Intravenous administration of puppy deciduous teeth stem cells in degenerative valve disease

Research (Published online: 16-12-2016)

15. Intravenous administration of puppy deciduous teeth stem cells in degenerative valve disease - Soontaree Petchdee and Sarunya Sompeewong

Veterinary World, 9(12): 1429-1434

   doi: 10.14202/vetworld.2016.1429-1434

Abstract

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Aim: The objective of this study is to investigate the improvement of heart function in dogs with chronic valvular heart disease after puppy deciduous teeth stem cells (pDSCs) administration.

Materials and Methods: 20 client-owned dogs with degenerative valvular heart disease underwent multiple intravenous injections of allogeneic pDSCs. Dogs were randomly assigned to two groups: (i) Control group (n=10) with standard treatment for heart failure and (ii) group with standard treatment and multiple administrations of pDSCs (n=10). Electrocardiography, complete transthoracic echocardiography, thoracic radiography, and blood pressure were recorded before and after pDSCs injections for 15, 30 and 60 days.

Results: Post pDSCs injection showed measurable improvement in left ventricular ejection fraction, American College of Veterinary Internal Medicine (ACVIM) functional class significantly improved and improved quality of life scores were observed. In the control group, there were no significant enhancements in heart function or ACVIM class.

Conclusions: This finding suggests that pDSCs could be a supplement for valvular heart disease treatment.

Keywords: dental pulp stem cells, dogs, heart failure.

Molecular characterization of field infectious bursal disease virus isolates from Nigeria

Research (Published online: 15-12-2016)

14. Molecular characterization of field infectious bursal disease virus isolates from Nigeria - Ijeoma O. Nwagbo, Ismaila Shittu, Chika I. Nwosuh, George O. Ezeifeka, Frederick J. C. Odibo, Linda O. Michel and Daral J. Jackwood

Veterinary World, 9(12): 1420-1428

   doi: 10.14202/vetworld.2016.1420-1428

Abstract

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Aim: To characterize field isolates of infectious bursal disease virus (IBDV) from outbreaks in nine states in Nigeria through reverse transcription polymerase chain reaction (RT-PCR) and sequence analysis of portions of the VP2 and VP1 genes and to determine the presence or absence of reassortant viruses.

Materials and Methods: A total of 377 bursa samples were collected from 201 suspected IBD outbreaks during 2009 to 2014 from nine states in Nigeria. Samples were subjected to RT-PCR using VP2 and VP1 gene specific primers, and the resulting PCR products were sequenced.

Results: A total of 143 samples were positive for IBDV by RT-PCR. These assays amplified a 743 bp fragment from nt 701 to 1444 in the IBDV VP2 hypervariable region (hvVP2) of segment A and a 722 bp fragment from nt 168 to 889 in the VP1 gene of segment B. RT-PCR products were sequenced, aligned and compared with reference IBDV sequences obtained from GenBank. All but one hvVP2 sequence showed similarity to very virulent IBDV (vvIBDV) reference strains, yet only 3 of the VP1 67 VP1 sequences showed similarity to the VP1 gene of vvIBDV. Phylogenetic analysis revealed a new lineage of Nigerian reassortant IBDV strains.

Conclusion: Phylogenetic analysis of partial sequences of genome segment A and B of IBDV in Nigeria confirmed the existence of vvIBDV in Nigeria. In addition, we noted the existence of reassortant IBDV strains with novel triplet amino acid motifs at positions 145, 146 and 147 in the reassorted Nigerian IBDV.

Keywords: infectious bursal disease virus, Nigeria, novel, reassortant, very virulent.

Evidence of hemolysis in pigs infected with highly virulent African swine fever virus

Research (Published online: 14-12-2016)

13. Evidence of hemolysis in pigs infected with highly virulent African swine fever virus - Zaven Karalyan, Hovakim Zakaryan, Elina Arakelova, Violeta Aivazyan, Marina Tatoyan, Armen Kotsinyan, Roza Izmailyan and Elena Karalova

Veterinary World, 9(12): 1413-1419

   doi: 10.14202/vetworld.2016.1413-1419

Abstract

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Aim: The research was conducted to understand more profoundly the pathogenetic aspects of the acute form of the African swine fever (ASF).

Materials and Methods: A total of 10 pigs were inoculated with ASF virus (ASFV) (genotype II) in the study of the red blood cells (RBCs), blood and urine biochemistry in the dynamics of disease.

Results: The major hematological differences observed in ASFV infected pigs were that the mean corpuscular volume, mean corpuscular hemoglobin, and hematocrits were significantly decreased compared to controls, and the levels of erythropoietin were significantly increased. Also were detected the trends of decrease in RBC count at terminal stages of ASF. Analysis of blood biochemistry revealed that during ASF development, besides bilirubinemia significantly elevated levels of lactate dehydrogenase, and aspartate aminotransferase were detected. Analysis of urine biochemistry revealed the presence of bilirubinuria, proteinuria during ASF development. Proteinuria, especially at late stages of the disease reflects a severe kidney damage possible glomerulonefritis.

Conclusion: The results of this study indicate the characteristics of developing hemolytic anemia observed in acute ASF (genotype II).

Keywords: African swine fever virus, bilirubinemia, bilirubinuria, erythropoietin, hemolysis, proteinuria.

Immunotoxic effect of thiamethoxam in immunized mice with Brucella abortus cultural filtrate antigen

Research (Published online: 13-12-2016)

12. Immunotoxic effect of thiamethoxam in immunized mice with Brucella abortus cultural filtrate antigen - L. H. Salema, M. J. Alwan and Afaf Abdulrahman Yousif

Veterinary World, 9(12): 1407-1412

   doi: 10.14202/vetworld.2016.1407-1412

Abstract

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Aim: This study was planned for determination the toxic effect of thiamethoxam (TMX) in immunized mice with Brucella abortus culture filtrate antigen (CFBAgs) (as a vaccine) and its role of TMX on decrease activity of B. abortus antigen on eliciting of humoral and cellular immunity.

Materials and Methods: To achieve these goals 60 female mice were used, 7-8 weeks age, they were divided equally into three groups (20 in each group) and treated as follows: 1st group: Mice were immunized with CFBAgs intraperitoneally in two doses, 2 weeks intervals with (protein concentration 2 mg\ml), 2nd group: Mice immunized as in the 1st group and was administrated orally with 1/10 lethal dose 50% of TMX (83.7 mg/kg B.W.) for 4 weeks daily, 3rd group was administrated orally with 0.3 ml normal saline served as a control group. At day 28 post immunization (PI) delayed type hypersensitivity (skin test) was done, and serum samples were collected at day 30 (PI) for detection of passive hemagglutination test (PHA); interferon gamma (IFN-γ) which was done by enzyme-linked immunosorbent assay test in addition to phagocytes assay.

Results: The results of skin test post injection with soluble antigen of B. abortus intradermally showed a high significantly mean values at p≤0.05 of footpad skin thickness in the 1st group of mice which recorded (0.51±0.002 mm) as compared with the 2nd group of mice which showed (0.08±0.002 mm) after 24 h; the mean values of skin thickness were declined in the 1st mice (0.46±0.002) and 2nd mice (0.070±0.001) at 48 h; control group showed a negative results. These results were agreed with results of serum levels of IFN-γ (pg/ml) that showed that a significant increase the vaccinated 1st group (406.36±1.52), than those values in the 2nd group (151.61±0.89) and negative result in 3rd group (46.47±0.60), in addition to results of PHA test which showed a significant increase in antibody titer in the 1st group (139±12.16) with low level of serum antibody in the 2nd group (7.66±0.33). Phagocytic ratio results in the 1st group showed an increase to reach (18.55±0.44) than a ratio in the 2nd group (13.24±0.32) and the control group (5.46±0.25).

Conclusion: It was concluded that TMX induced suppression of humoral and cellular immune responses in immunized mice with CFBAgs.

Keywords: Brucella abortus, enzyme-linked immunosorbent assay test, interferon gamma, phagocyte assay, skin test, thiamethoxam.