Isolation and characterization of avian coronavirus from healthy Eclectus parrots (Eclectus roratus) from Indonesia

Research (Published online: 19-11-2019)

16. Isolation and characterization of avian coronavirus from healthy Eclectus parrots (Eclectus roratus) from Indonesia

G. K. Suryaman, R. D. Soejoedono, A. Setiyono, O. N. Poetri and E. Handharyani

Veterinary World, 12(11): 1797-1805

ABSTRACT

Background and Aim: Avian coronavirus has a wide range of hosts, from chickens and turkeys to wild birds. This virus causes an economically and, possibly, environmentally, important loss in the poultry industry. Therefore, research into the avian coronavirus in various species of birds is required. The Eclectus parrot (Eclectus roratus) is an endemic bird to Indonesia and Northern Australia and often kept as pets. At present, there has been limited information about avian coronavirus infection among birds. This study aimed to determine the presence of and to characterize avian coronavirus isolated from Eclectus parrots in Indonesia.

Materials and Methods: Cloacal swab samples were taken from 10 healthy Eclectus parrots (E. roratus). Each isolate was propagated into specific pathogen-free embryonated chicken eggs. The presence of avian coronavirus was determined using three sets of primers targeting the 3' untranslated region (3'-UTR) of avian coronavirus (UTR41+/11–), the N gene of the infectious bronchitis virus (IBVN+/–), and the S1 gene of the IBV (XCE2+/XCE2–). The infectious bronchitis vaccine strain H120 was used as a positive control. Resulting positive bands were sequenced for the S1 gene.

Results: None of the isolates was positive for the 3'-UTR, four isolates were positive for the N gene of infectious bronchitis, and two isolates were positive for the S1 gene of the IBV. However, only one isolate (parrot/Indonesia/BX9/16) was sequenced for the partial S1 gene with primers XCE2+/XCE2–. The partial nucleotide sequence of this isolate showed 100% homology with the IBV GI-13 lineage, specifically with a field isolate of the 4/91 variant 1 Israel and the 4/91 vaccine on the hypervariable region 3 site of the S1 gene.

Conclusion: An IB-like avian coronavirus was isolated from healthy Eclectus parrots. Our results indicate that IBV has a wide range of hosts, which prompt the need to understand the interspecies connection of this virus better.

Keywords: Avian coronavirus, Eclectus parrot, Infectious bronchitis.

Endotheliotropic herpesvirus infection in Asian elephants (Elephas maximus) of Assam, India

Research (Published online: 18-11-2019)

15. Endotheliotropic herpesvirus infection in Asian elephants (Elephas maximus) of Assam, India

G. Mahato, K. K. Sarma, D. C. Pathak, N. N. Barman, P. Gogoi, M. Dutta and P. Basumatary

Veterinary World, 12(11): 1790-1796

ABSTRACT

Background and Aim: Elephant endotheliotropic herpesvirus (EEHV) is an emerging disease of elephant. Therefore, a study was conducted to know the actual status of the disease in Assam State of India.

Materials and Methods: A total of 289 Asian elephants of Assam were screened during 2 years of study from April 2017 to March 2019. The clinical symptoms of diseased as well as gross and histopathological changes of dead elephants were recorded for the diagnosis of the disease. Virus involved in the occurrence of the disease was confirmed by polymerase chain reaction (PCR).

Results: In the present study, a total of three elephant calves out of 22 were found positive to EEHV1A. On the other hand, three adult asymptomatic elephants were also found positive for EEHV1 on screening 267 captive Asian elephants of Assam. The amplified PCR product showed band size of 520, 600, and 930 bp. The PCR amplified product with size 600 bp had shown the gene sequence for EEHV1U77/HEL. Gross lesions include congested blood vessels of the liver and intestinal mucosa, foci of petechiae in the spleen, and heart and focal ulceration in the dorsal surface of the tongue. Microscopically, the kidneys showed intertubular edema and focal areas of degeneration associated with coagulative necrosis of the tubular epithelium. The liver showed hydropic degeneration and fatty changes of the hepatocytes. There was a massive proliferation of fibroblasts in the interlobular spaces which penetrated the necrosed areas of the hepatic lobules.

Conclusion: A total of three wild rescued elephant calves and three asymptomatic adults were found positive for EEHV1A during the 2 years of study. The PCR amplified product with size 600 bp had shown the gene sequence for EEHV1U77/HEL.

Keywords: amplicons, amplification, elephant endotheliotropic herpesvirus, phylogenetic, polymerase chain reaction.

Addition of L-arginine in skim milk extender maintains goat spermatozoa quality in chilled temperature for five days

Research (Published online: 15-11-2019)

14. Addition of L-arginine in skim milk extender maintains goat spermatozoa quality in chilled temperature for five days

Suherni Susilowati, Indah Norma Triana, Wurlina Wurlina, Arimbi Arimbi, Pudji Srianto and Imam Mustofa

Veterinary World, 12(11): 1784-1789

ABSTRACT

Aim: The purpose of this study was to determine the benefits of L-arginine addition in skim milk extender to maintain the quality of goat spermatozoa in chilled storage.

Materials and Methods: A total of 18 ejaculates from three healthy goats with weight and age of 45 kg and 4-5 years, respectively, were divided into three groups. The control group contained goat semen diluted in a skim milk extender without L-arginine; Treatment I and Treatment II contained goat semen diluted in a skim milk extender with added L-arginine 4 and 6 mM, respectively. These three groups were chilled at 5°C and evaluated daily for 5 days. Observed variables were viability, motility, intact plasma membrane (IPM), malondialdehyde (MDA) level, necrosis, and apoptosis of spermatozoa.

Results: The addition of L-arginine 4 mM was the best treatment in maintaining viability, motility, and IPM and a decreased MDA level, percentage of necrosis, and apoptosis of goat spermatozoa. An ejaculate in this extender can be divided into 37 doses for intracervical insemination in <1 ml volume with 125 million motile spermatozoa.

Conclusion: Goat semen retained its quality when kept for 5 days in chilled storage by adding L-arginine in skim milk extender.

Keywords: apoptosis, goat spermatozoa, L-arginine, malondialdehyde level, necrosis.

Pregnancy rates in hair sheep after Ovsynch synchronization and a combined intracervical fixed-time artificial insemination and 10-day mating period

Research (Published online: 15-11-2019)

13. Pregnancy rates in hair sheep after Ovsynch synchronization and a combined intracervical fixed-time artificial insemination and 10-day mating period

D. A. Vallejo, J. D. Londoño, Y. A. Yepes, V. Tamayo, A. F. Mejia and J. G. Maldonado

Veterinary World, 12(11): 1779-1783

ABSTRACT

Aim: This study aimed to evaluate the pregnancy rates in hair ewes using an Ovsynch synchronization protocol under a breeding system that combines fixed-time insemination plus a 10-day mating period as an alternative.

Materials and Methods: Through an experimental study (n=27), ewes were randomly located into one of three treatments: (1) Pre-synch (n=9): Prostaglandin F2α (PGF2α)+Gonadotropin-releasing hormone (GnRH)+PGF2α+GnRH; (2) Ovsynch (n=9): GnRH+PGF2α+GnRH; and (3) control: Ewes bred by natural mating (NM) (n=9). Ewes were fixed-time inseminated (fixed-time artificial insemination [FTAI]) with fresh semen, collected just before the insemination time through vaginoscopy at 16 h after the second GnRH (gonadorelin) injection. Each experimental group was placed separately during 15 days and, after this time, fertile rams were allowed back with ewes for a 10-day mating period. Control group ewes remained with the rest of the herd suitable for breeding and were bred under NM. Pregnancy diagnosis was performed by ultrasound at 28-, 56-, and 84-day post-breeding to differentiate between FTAI and NM pregnancies. Total (FTAI±NM) pregnancy rates at 56-day post-breeding were used to compared Pre-synch, Ovsynch, and control. For this purpose, two-tailed proportions comparison z-test was used with a 95% confidence level, for testing as the null hypothesis whether two proportions were equal.

Results: Pregnancy rates were higher in control ewes (66.4%) than FTAI (46.6%). When pregnancy rates after a 10-day mating period (40%) were added, the final rate (86.6%) was significantly (p<0.05) higher in Ovsynch-based protocols. The pregnancy rate was significantly lower in FTAI ewes compared to FTAI +10-day mating group (p<0.05). The overall pregnancy rate was 88.0, 85.7, and 67.0 (p>0.05) for Pre-synch, Ovsynch, and control ewes, respectively.

Conclusion: These results provide evidence on the benefits of combined FTAI protocols for improving the reproductive efficiency of sheep.

Keywords: ewes, fixed-time artificial insemination, Ovsynch, pre-synch.

Evaluation of effects of a novel probiotic feed supplement on the quality of broiler meat

Research (Published online: 13-11-2019)

12. Evaluation of effects of a novel probiotic feed supplement on the quality of broiler meat

Ujang Suryadi, Yudhi Ratna Nugraheni, Anang Febri Prasetyo and Aan Awaludin

Veterinary World, 12(11): 1775-1778

ABSTRACT

Background and Aim: A local microorganism-based probiotic has been developed as an alternative to using antibiotic growth promoter and its effect on broiler meat quality has been studied, when supplemented with poultry feed at different concentrations. This study aimed to understand the effect of local microorganism-based probiotic sourced from cattle rumen and chicken intestine and added as feed supplement at different concentrations on broiler meat quality.

Materials and Methods: The local microorganism-based probiotic made from cattle rumen and chicken intestine contained Lactobacillus spp., Bifidobacterium spp., Streptococcus spp., and Bacillus spp. The experiments followed a completely randomized design. Treatments in the study were: P0, i.e., control (without probiotic), P1 (probiotic administered at 5 ml/kg feed), P2 (probiotic administered at 10 ml/kg feed), and P3 (probiotic administered at 15 ml/kg feed). Each treatment was repeated 5 times. Parameters examined in this study were pH, meat tenderness, fat content, and meat protein content.

Results: Based on a total of 200 chickens, the percentage of meat protein content in treatments P1, P2, and P3 showed an increase of 19.34%, 19.42%, and 19.64%, respectively, when compared with P0 that showed a protein content increase of 19.14%. The fat content of meat for P1, P2, and P3 was 21.54%, 21.46%, and 21.30%, respectively, which was less than the value for P0 (21.69%). The treatments did not significantly affect pH or meat tenderness when compared with the control. The usage of this novel probiotic as a feed supplement resulted in an increase in meat protein content and a decrease in fat content.

Conclusion: This study indicates that using the local microorganism-based probiotic sourced from cattle rumen and chicken intestine to supplement poultry feed did not have a significantly different effect (p>0.05) on meat pH; however, it had a significantly different (p<0.05) on protein and fat content of broiler meat.

Keywords: broiler, chicken, meat, microorganism, probiotic.

Application of immunofluorescence assay and nested polymerase chain reaction for query fever diagnosis in animal handlers of Puducherry, South India, and phylogenetic analysis based on IS1111 repetitive gene element

Research (Published online: 13-11-2019)

11. Application of immunofluorescence assay and nested polymerase chain reaction for query fever diagnosis in animal handlers of Puducherry, South India, and phylogenetic analysis based on IS1111 repetitive gene element

Jothimani Pradeep, Selvaraj Stephen, Balakrishnan Sangeetha, Prabakar Xavier Antony, S. Amsaveni and Pratheesh Pooja

Veterinary World, 12(11): 1769-1774

ABSTRACT

Background and Aim: Diagnosis of query fever (QF) is mostly done on the basis of serological/molecular tests, due to the stringent requirement of biosafety level-3 containment facilities for isolating Coxiella burnetii in culture. QF is an important zoonosis and is considered to be an occupational hazard to livestock handlers. This report describes our study on the serological as well as molecular evidence of QF in animal handlers from Puducherry and surrounding Tamil Nadu, from where, to the best of our knowledge, no such reports are available so far.

Materials and Methods: Seventy-five animal handlers were recruited, comprising veterinarians, slaughterhouse workers, butchers, and animal attendants of various government veterinary clinics from Puducherry and surrounding areas of Tamil Nadu state. QF serology was performed to identify Phase I and Phase II immunoglobulin G antibodies to C. burnetii. Nested polymerase chain reaction (N-PCR) was carried out to detect C. burnetii DNA in buffy coat samples by targeting IS1111 gene element. N-PCR-positive samples were sequenced and phylogenetic analysis was performed using MEGA software version 10.0.

Results: A total of 21 animal handlers (28.1%) were positive for either serology or PCR. PCR alone was positive in 10 (13.4%), only serology was positive in 8 (10.7%), and both serology and PCR were positive in three samples (4.0%). GenBank accession numbers were obtained for 13 N-PCR-positive samples (MG548608-MG548620). Six of our study sequences showed close similarity with the reference isolates from Bengaluru, Colombia, Brazil, France, and Iran.

Conclusion: A significant percentage of QF positivity in animal handlers of this part of South India, Puducherry, warrants a prospective study with follow-up of a large number of this occupational group.

Keywords: Coxiella burnetii, immunofluorescence assay immunoglobulin G, Q fever in Puducherry, query fever nested polymerase chain reaction, zoonosis.

Molecular detection and typing of methicillin-resistant Staphylococcus aureus and methicillin-resistant coagulase-negative staphylococci isolated from cattle, animal handlers, and their environment from Karnataka, Southern Province of India

Research (Published online: 11-11-2019)

10. Molecular detection and typing of methicillin-resistant Staphylococcus aureus and methicillin-resistant coagulase-negative staphylococci isolated from cattle, animal handlers, and their environment from Karnataka, Southern Province of India

Nimita Venugopal, Susweta Mitra, Rituparna Tewari, Feroze Ganaie, Rajeswari Shome, Habibur Rahman and Bibek R. Shome

Veterinary World, 12(11): 1760-1768

ABSTRACT

Background and Aim: Methicillin-resistant staphylococci are among the emerging pathogens which have become a threat to both human and animal health. The present investigation intended to examine the occurrence and the molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant coagulase-negative staphylococci (MRCoNS) recovered from cattle, its handlers, and their environment.

Materials and Methods: A total of 666 specimens were subjected to culture method and genus-specific polymerase chain reaction (PCR) for the identification of Staphylococcus. Methicillin resistance was substantiated by PCR identification of mecA and mecC resistance determinants. Species-specific identification of mecA positive isolates was conducted by multiplex PCR. The unidentified species were deciphered by 16S rRNA gene sequencing approach. The mecA positive isolates were further characterized by staphylococcal cassette chromosome mec (SCCmec) typing and multilocus sequence typing (MLST).

Results: Duplex PCR identified 728 Staphylococcus isolates, of which 66 (9%) were positive for mecA gene. MRSA constituted 24% of the total mecA positive isolates. Among MRCoNS, Staphylococcus epidermidis (42%), and Staphylococcus haemolyticus (11%) were the most common species identified. Overall, 47% of the mecA positive isolates belonged to SCCmec type V. MLST analysis showed eight different sequence types (STs) among MRSA isolates of which five were novel STs. Among methicillin-resistant S. epidermidis, 19 different STs were found, of which nine novel STs were detected.

Conclusion: The increase in the prevalence of mecA positive staphylococci, especially MRCoNS in cattle is a great concern in view of their transmission potential. Hence, continuous monitoring and molecular characterization of methicillin-resistant staphylococci should be elucidated in human and animal sectors so as to prevent the spread of these resistant pathogens.

Keywords: mecA/mecC gene, methicillin resistance, methicillin-resistant coagulase-negative staphylococci, methicillin-resistant Staphylococcus aureus.