Immunopathological immunohistochemical study of low pathogenic avian influenza virus H5N1 infection in lovebirds (Agapornis spp.) in Indonesia

Research (Published online: 26-09-2019)

18. Immunopathological immunohistochemical study of low pathogenic avian influenza virus H5N1 infection in lovebirds (Agapornis spp.) in Indonesia

Zulfikhar Zulfikhar, Raden Wasito and Hastari Wuryastuti

Veterinary World, 12(9): 1472-1477

ABSTRACT

Background and Aim: To date, persistent infection with low pathogenic avian influenza virus (AIV) subtype H5N1 (LPAI H5N1) in chickens is widespread in Indonesia. Commercially available ornamental birds on markets play an important role in triggering the occurrence of AIV outbreaks. Thus, the presence of AIV in ornamental birds such as lovebirds needs to be closely monitored. Here, the presence of LPAI H5N1 was investigated in lovebirds with no clinical signs that, although apparently healthy, can be a continuous source of AIV transmission to other sensitive birds such as chickens and even humans.

Materials and Methods: In the present study, 30 lovebirds were necropsied. The immunopathology of the lungs, which had hemorrhages, was immunohistochemically studied using the streptavidin-biotin (SB) method to stain for LPAI H5N1. Polyclonal antibodies against the nucleoprotein or monoclonal antibodies against the hemagglutinin of the AIV subtype H5N1 were used to identify these antigens in lung tissue samples.

Results: LPAI H5N1 was detected in the lungs of all lovebirds according to the brownish colored deposits in the histological samples. The highest intensity of the SB staining was found in the lumina of vascular capillaries and the cell membranes of air capillaries. The discovery of LPAI H5N1 in lovebirds increases the environmental health risk and raises the possibility of exposure to AIV. Thus, it is very important to improve the mechanisms that control the traffic of ornamental birds between regions and islands, as well as the AIV vaccination strategies related to the prevention, control, and eradication of AIV in Indonesia, and these procedures should be reevaluated.

Conclusion: The present study proves that lovebirds are infected with LPAI subtype H5N1. Even if ornamental birds such as lovebirds do not show any clinical signs, they are likely to be a persistently infected with LPAI H5N1. Therefore, ornamental birds might be a continuous source of LPAI H5N1 infections in other sensitive birds, including poultry, and may also be responsible for virus transmission to humans.

Keywords: avian influenza virus H5N1, immunohistochemistry, Indonesia, lovebird, lung, streptavidin-biotin.

Detection of torque teno sus virus infection in Indian pigs

Research (Published online: 25-09-2019)

17. Detection of torque teno sus virus infection in Indian pigs

Vinutha Subramanyam, Divakar Hemadri, Shashidhara Phani Kashyap, Jagadish Hiremath, Nagendra Nath Barman, Esther Lalzoliani Ralte, Sharanagouda S. Patil, Kuralayanapalya P. Suresh and Habibur Rahaman

Veterinary World, 12(9): 1467-1471

ABSTRACT

Background and Aim: Torque teno viruses (TTVs) are circular, single-stranded DNA viruses, which infect a wide range of animals including livestock and companion animals. Swine TTVs (torque teno sus viruses [TTSuVs]) are thought to act as a primary or coinfecting pathogen in pathological conditions such as porcine dermatitis and nephropathy syndrome and post-weaning multisystemic wasting syndrome. So far, the presence of the virus has not been reported in India. Considering that TTSuVs have the potential to cross the species barrier into humans and that pork consumption is common in North-Eastern states of India, the current study aims to investigate the presence of TTSuV in the Indian pig population.

Materials and Methods: A total of 416 samples were collected during 2014-2018, from both apparently healthy pigs and also from pigs suspected of having died from classical swine fever and/or porcine reproductive and respiratory syndrome. These samples were screened for TTSuV infection by polymerase chain reaction (PCR) and DNA sequencing techniques.

Results: The presence of the virus was confirmed in 110 samples from 12 different states of India. Phylogenetic analysis of the nucleotide sequences obtained from the PCR products indicated the presence of viruses of both Iotatorquevirus and Kappatorquevirus genera in India.

Conclusion: The study is the first report on the presence of TTSuVs in India and highlights the circulation of both genera of the virus in the country.

Keywords: detection, India, phylogeny, polymerase chain reaction, torque teno virus.

The distribution of serotonergic nerve on the hippocampus of the fruit bats (Rousettus amplexicaudatus)

Research (Published online: 25-09-2019)

16. The distribution of serotonergic nerve on the hippocampus of the fruit bats (Rousettus amplexicaudatus)

Vivin Wirawati, Nourrisma D. A. Widiati, Geraldus Gunawan, Golda R. Saragih, Puspa Hening and Hevi Wihadmadyatami

Veterinary World, 12(9): 1460-1466

ABSTRACT

Background: Rousettus amplexicaudatus is one of the old world fruit bats, commonly known in Javanese word as codot belongs to Order Chiroptera and suborder Megachiroptera. R. amplexicaudatus possessed an outstanding visual ability, which helps in the process of searching for food. Those activities process and save within the brain involving the learning and memory activities. Learning and memory activities are centered around the hippocampus with the help of serotonergic nerve.

Aim: This study conducted to identify the morphology and distribution of serotonergic nerve (5-hydroxytryptamine) in the hippocampus of R. amplexicaudatus, which related to the function of serotonin within the learning and memory process.

Materials and Methods: Five R. amplexicaudatus bats were brought from Gunung Kidul cave, Yogyakarta, Java Island, Indonesia. The bats were anesthetized using ketamine and xylazine. R. amplexicaudatus then perfused intracardially. The brain samples were collected processed into paraffin block, and a further cut in 8 μm thickness serially. The tissue slides were stained using hematoxylin-eosin, cresyl echt violet, and immunohistochemistry with rabbit's anti-serotonin (1:200). The results observed and analyzed quantitatively and qualitatively using image J (https://imagej.nih.gov/ij/) (Bethesda, Maryland, USA) and GraphPad Prism 7 (La Jolla, CA, USA).

Results: The hippocampus of R. amplexicaudatus composed of the dentate gyrus (DG), hippocampus proper (cornu ammonis 3 [CA3], and CA1 as the main area for learning memory), and subiculum. On the DG serotonin immunoreactive cells found within the granular layer (132±35.03 cells/mm2), polymorphic stratum (86.33±11.23 cells/mm2), and molecular layer (93±1 cells/mm2). Meanwhile, on CA3 area, the immunoreactive cells for serotonin found in each stratum. The number of immunoreactive cells on each stratum from highest to the lowest are stratum pyramidal 123.33±15.88 cell/mm2, stratum molecular 63±13.11 cell/mm2, stratum lucidum 62.67±8.08 cell/mm2, stratum radiatum 55.33±510.21 cell/mm2, stratum oriens 48±3.46 cell/mm2, and stratum alveus 28.67±2.52 cell/mm2. In addition, in CA1 also hampers the immunoreactive cells in the pyramidal stratum, molecular, lucidum, oriens, radiatum, and alveus layer, respectively, of each 91±27.40 cell/mm2, 60.33±20.65 cell/mm2, 53.67±4.51 cell/mm2 44.33±10.40 cell/mm2, 41.33±5.51 cell/mm2, and 27±4 cell/mm2.

Conclusion: Taking together the distribution of serotonin-immunoreactive cells in the hippocampus of R. amplexicaudatus mostly found on CA3 followed by CA 1 and DG.

Keywords: hippocampus, immunohistochemistry, learning and memory, Rousettus amplexicaudatus, serotonin.

Detection of Leptospira spp. using polymerase chain reaction technique from kidney of Rattus norvegicus from Grenada, West Indies

Research (Published online: 24-09-2019)

12. Detection of Leptospira spp. using polymerase chain reaction technique from kidney of Rattus norvegicus from Grenada, West Indies

Bhumika Sharma, Katelyn Thille, Nia Rametta and Ravindra Sharma

International Journal of One Health, 5: 81-85

ABSTRACT

Aim: This study aimed to find out the prevalence of active infection of Leptospira spp. in Rattus norvegicus from Grenada, West Indies, through polymerase chain reaction (PCR).

Materials and Methods: One hundred and forty-nine rats were trapped, anesthetized and their kidneys collected aseptically. DNA was extracted from the kidney tissue of each rat. PCR was performed targeting LipL32 gene. Eighteen PCR-positive amplicons for LipL32 gene segment were purified and sent for direct sequencing to the sequencing facility of MCLAB (South San Francisco, USA). Results of sequencing were read and interpreted. The prevalence of Leptospira spp. in relation to sex and age was also recorded.

Results: All amplified sequences were compared to the sequences present in GenBank using basic local alignment search tool (BLAST) from the online website National Center for Biotechnology Information, the results revealed that six samples had similarity to Leptospira interrogans strain 1399/2016 and eight samples had similarity with Leptospira borgpetersenii serovar Hardjo-bovis strain L49. Of 149 kidney samples, only 14 were positive for Leptospira spp. by PCR giving an incidence of 9.3%. There was no significant difference found in relation to sex and age.

Conclusion: This is the first report confirming active infection of Leptospira spp. in Rattus norvegicus in Grenada using PCR. The presence of active infection in rats can be considered as high risk for humans. Further research to understand the epidemiology of leptospirosis in Grenada is suggested.

Keywords: Rattus norvegicus, Grenada, kidney, Leptospira spp., polymerase chain reaction.

Prevalence and species identification of Cryptosporidium spp. in the newborn dairy calves from Muang District, Khon Kaen Province, Thailand

Research (Published online: 23-09-2019)

15. Prevalence and species identification of Cryptosporidium spp. in the newborn dairy calves from Muang District, Khon Kaen Province, Thailand

Phennarin Doungmala, Patchara Phuektes, Weerapol Taweenan, Somboon Sangmaneedet and Ornampai Japa

Veterinary World, 12(9): 1454-1459

ABSTRACT

Aim: This study aims to determine the prevalence of Cryptosporidium spp. infection and to identify the species of Cryptosporidium spp. in newborn dairy calves between December 2016 and March 2017 in Muang District, Khon Kaen Province, Thailand.

Materials and Methods: A total of 200 fecal samples from newborn dairy calves of the ages 1 day up to 28 days were collected and the presence of Cryptosporidium oocysts was examined microscopically using the modified Kinyoun's acid-fast staining technique. Then, Cryptosporidium species were identified using nested polymerase chain reaction amplification of 18S rRNA gene and sequencing.

Results: The modified Kinyoun's acid-fast staining revealed the presence of Cryptosporidium oocysts in 51% (102/200). Sequence analysis of the 18S rRNA gene identified two species, namely, Cryptosporidium bovis (n=11) and Cryptosporidium ryanae (n=11) and one isolated strain could not be identified.

Conclusion: This study indicated that newborn dairy calves aging up to 4 weeks were highly infected with Cryptosporidium spp., and the infection mostly occurred in diarrheic dairy calves. This is the first report of Cryptosporidium in dairy calves in Khon Kaen Province and the results provide baseline information for further studies and control of Cryptosporidium infection in dairy calves in the study area.

Keywords: Cryptosporidium, Khon Kaen Province, nested polymerase chain reaction, newborn dairy calves, prevalence.

Modification on acute myocardial infarction model through left anterior descending coronary artery ligation: An experimental study on rats (Rattus norvegicus) using readily available materials

Research (Published online: 21-09-2019)

14. Modification on acute myocardial infarction model through left anterior descending coronary artery ligation: An experimental study on rats (Rattus norvegicus) using readily available materials

Johanes Nugroho, Wiwik Misaco Yuniarti, Ardyan Wardhana and Cornelia Ghea

Veterinary World, 12(9): 1448-1453

ABSTRACT

Background and Aim: Several difficulties are involved in creating models for myocardial infarction (MI) in animals, such as low survival rates after acute MI, complicated techniques in creating animal models, complexities in confirming acute MI incidence, and complex surgical tools needed in the process. This study aimed to develop an animal model for acute MI using Wistar rats utilizing simple instruments that are readily available in standard animal laboratories.

Materials and Methods: We induced MI in 48 Wistar rats using the left anterior descending coronary artery ligation modification technique without tracheal incision and ventilator. This ligation technique was performed 1-2 mm distal to the left atrial appendage. MI occurrence was evaluated using heart enzyme parameters 24 h post-ligation and histological studies of the infarcted area 6 weeks after the ligation. Rats were divided into the coronary artery ligation group and sham group.

Results: Of the 48 rats, 24 (50%) died within 24 h post-ligation, but no further deaths occurred in the next follow-up period of 6 weeks. The average infarct size in six rats within 24 h of ligation was 35%±5.7%. The serum glutamic oxaloacetic transaminase level of the group treated with coronary artery ligation was statistically significantly higher than that of the sham group (p=0.000).

Conclusion: We developed an MI rat model with consistent infarction size, in which the long-term death of rats was not observed. Our ligation technique for an MI rat model can be a reference for experimental settings without ventilators for small animals.

Keywords: left anterior descending coronary artery ligation, myocardial infarction, rat model.

Assessing farmers' perspective on antibiotic usage and management practices in small-scale layer farms of Mymensingh district, Bangladesh

Research (Published online: 19-09-2019)

13. Assessing farmers' perspective on antibiotic usage and management practices in small-scale layer farms of Mymensingh district, Bangladesh

Jannatul Ferdous, Sabbya Sachi, Zakaria Al Noman, S. M. Azizul Karim Hussani, Yousuf Ali Sarker and Mahmudul Hasan Sikder

Veterinary World, 12(9): 1441-1447

Background and Aim: Indiscriminate and injudicious use of antibiotics in layer farms is a common practice of Bangladesh for the compensation of Poor management practices and ignorance. Despite this scenario, there is no published documentation on antibiotic usage pattern and farm management practices in layer farms. This study was undertaken to understand the farmers' perspective in small-scale layer farms regarding antibiotics usage and farm management.

Materials and Methods: A questionnaire survey was conducted in 120 small-scale layer farms of Mymensingh district during January-February 2017. We only considered farms in production. Data were analyzed on antibiotic usage, purpose, egg management, understanding of antibiotic residue, withdrawal period, and other issues.

Results: Among 120 farmers, about 94.16% of farmers are using antibiotics without respecting the withdrawal period. Only 39.1% of farmers possess knowledge of residues. In our surveyed farms, 91.83% of farmers are not practicing egg washing before supplying to the market and 52.67% of farmers are unaware of cleaning and disinfection of egg tray. Ten different types of antibiotics of seven classes have found in the survey. Most antibiotics are in the Watch (49%) and Reserve (8%) groups according to the WHO AWaRe categorization and 73% antibiotics are critically important for human medicine and are considered as last resort.

Conclusion: This study found that due to the lack of knowledge and poor management, farmers consider using antibiotics as the most effective practices to control disease and enhancement of egg production. These indiscriminate uses of antibiotics are responsible for antibiotic residual and resistance problem. Here, we also provide some suggestion and guidelines to improve management practices to minimize the emerging problems of antimicrobial resistance through small-scale layer farms.

Keywords: antibiotic usage, AWaRe categorization, layer farms, management practices.