Identification of uncultured bacteria from abscesses of exotic pet animals using broad-range nested 16S rRNA polymerase chain reaction and Sanger sequencing

Research (Published online: 09-10-2019)

6. Identification of uncultured bacteria from abscesses of exotic pet animals using broad-range nested 16S rRNA polymerase chain reaction and Sanger sequencing

T. Duangurai, J. Siengsanan-Lamont, C. Bumrungpun, G. Kaewmongkol, L. Areevijittrakul, T. Sirinarumitr, S. G. Fenwick and S. Kaewmongkol

Veterinary World, 12(10): 1546-1553

ABSTRACT

Background: The Sanger sequencing technique has been questioned and challenged by advanced high-throughput sequencing approaches. Sanger sequencing seems to be an obsolete technology. However, there are still research problems that could be answered using the Sanger sequencing technology. Fastidious obligate anaerobic bacteria are mostly associated with abscesses in animals. These bacteria are difficult to isolate from abscesses and are frequently excluded due to the bias of conventional bacterial culturing.

Aim: This study demonstrated the usefulness of a broad-range polymerase chain reaction (PCR) with Sanger sequencing to identify the majority population of bacteria in abscesses from exotic pet animals.

Materials and Methods: This study performed a pilot investigation of abscesses from 20 clinical cases (17 rabbits, 2 hedgehogs, and 1 sugar glider) using standard culture methods for both aerobes and anaerobes and broad-range nested PCR targeting the 16S rRNA gene followed by the Sanger sequencing technique.

Results: The standard culture and PCR techniques detected bacteria in 9 and 17 of 20 samples, respectively. From the 17 sequencings of the 16S rRNA, 10 PCR products were found to be closely related with obligate anaerobes including Bacteroides spp., Fusobacterium spp., Prevotella spp. Phylogenetic analysis using the rpoB gene revealed that the species for the Bacteroides was thetaiotaomicron and for the Fusobacterium was varium and nucleatum. However, the amplification of the rpoB gene for the Prevotella spp. was unsuccessful. Correlations between the standard culture and PCR techniques were found in 9 (6 positive and 3 negative samples) of 20 samples. Eleven samples were discordant between the standard culture and PCR techniques which were composed of eight samples negative by culture but positive by PCR and three samples had different bacteria by the culture and PCR techniques.

Conclusion: According to this study, broad-range PCR combined with Sanger sequencing might be useful for the detection of dominant anaerobic bacteria in abscesses that were overlooked based on conventional bacterial culture.

Keywords: anaerobic bacteria, abscesses, exotic pet animals, Sanger sequencing.

Potential of medicinal plants to treat dengue

Review (Published online: 08-10-2019)

13. Potential of medicinal plants to treat dengue

Dulanjalee Neelawala, Sanjaya Rajapakse and Wikum Widuranga Kumbukgolla

International Journal of One Health, 5: 86-91

ABSTRACT

Dengue is a major public health challenge worldwide, particularly in tropical areas. Nearly 390 million infections and 22,000 deaths occur every year. At present, there are no specific therapeutics available to treat dengue; however, possible treatment procedures are explained in the traditional medical systems (TMSs), such as Sri Lankan TMS, Indian Ayurvedic, Unani, and Siddha TMS. In these TMSs, medicinal plants have been used in several ways against dengue, such as virocides, larvicides, and mosquito repellents. Therefore, medicinal plants inherit biologically active compounds/lead compounds that are yet to be identified chemically and physiologically. Herein, we discuss the possible applications of crude plant extracts and isolated phytochemicals from medicinal plants such as quercetin, sulfated galactomannans, flavonoids, and glabranine in controlling dengue. Moreover, medicinal plant-based therapeutics can be safer, cost-effective, and non-toxic. Therefore, this paper reviews the medicinal plants that are used in TMSs to manage dengue, the phytochemicals they contain, and mode of action of these phytochemicals such as virocides, larvicides, and mosquito repellents.

Keywords: dengue, in silico, larvicides, phytochemicals, virocides.

Identification of Staphylococcus species isolated from preputium of Aceh cattle based on 16S rRNA gene sequences analysis

Research (Published online: 08-10-2019)

5. Identification of Staphylococcus species isolated from preputium of Aceh cattle based on 16S rRNA gene sequences analysis

Muhammad Hambal, Masda Admi, Safika Safika, Wahyu Eka Sari, Teuku Reza Ferasyi, Dasrul Dasrul, Ummu Balqis and Darmawi Darmawi

Veterinary World, 12(10): 1540-1545

ABSTRACT

Aim: This research aimed to identify Staphylococcus species isolated from preputial swabs of healthy Aceh cattle, based on 16S ribosomal RNA gene analysis.

Materials and Methods: The bacterium was isolated from preputial swabs of healthy Aceh cattle. The total DNA from the isolated bacteria was extracted using the Genomic DNA Mini Kit followed by polymerase chain reaction (PCR) amplification of the 16S rRNA gene. The product of PCR amplification was then sequenced and aligned to the known sequences in the GenBank database by multiple alignments and was also analyzed by bioinformatics software to construct a phylogenetic tree.

Results: The results revealed that the bacterial isolate 3A had genetically closed relation to Staphylococcus pasteuri with <97% maximum identity. Data derived from the phylogenetic tree revealed that the bacterial isolate 3A was also related to Staphylococcus warneri, yet, it shows a different evolutionary distance with the ancestors (S. pasteuri).

Conclusion: The results of this research suggested that the bacterium 3A, isolated from preputial swabs of healthy Aceh cattle, is a Staphylococcus species.

Keywords: 16S rRNA gene, Aceh cattle, phylogenetic tree, polymerase chain reaction, Staphylococcus pasteuri.

Antioxidative activity and protein profile of skim milk of Gaddi goats and hill cattle of North West Himalayan region

Research (Published online: 05-10-2019)

4. Antioxidative activity and protein profile of skim milk of Gaddi goats and hill cattle of North West Himalayan region

Vinesh Sharma, Birbal Singh, Rinku Sharma, Jyoti B. Dhar, Neelam Sharma and Gorakh Mal

Veterinary World, 12(10): 1535-1539

ABSTRACT

Aim: This study was aimed at evaluation of antioxidative activity, protein profile, and vitamins content of milk of Gaddi goats, local non-Gaddi goats, hill cattle, and Jersey crossbred cattle.

Materials and Methods: Total phenol, antioxidant activity measured as 2, 2-diphenyl- 1-picrylhydrazyl radical scavenging capacity, total protein, and vitamins were estimated in milk samples by spectrophotometric methods. Milk protein profiles were studied by sulfate-polyacrylamide gel electrophoresis.

Results: Total phenol, antioxidant activity, and total protein were higher in indigenous hill cattle skim milk. Average protein content in raw skimmed milk was 1.33±0.01, 1.03±0.02, 0.76±0.05, and 0.81±0.01%, in indigenous hill cattle, Jersey crossbred cattle, non-Gaddi goat, and Gaddi goat, respectively. Three proteins of 19.01, 22.08, and 32.96 kDa were observed in Gaddi goat, but not in non-Gaddi goat skim milk. Furthermore, the above proteins were absent in cattle skim milk. Two proteins of 15.56 and 25.06 kDa were found in local hill and crossbred cattle skimmed milk, but were absent in goat skimmed milk. Vitamin C content was the lowest in Gaddi goat milk and the highest in Jersey crossbred cattle milk.

Conclusion: It is envisaged that bioactive metabolites in the milk of Gaddi goats and hill cattle might offer anti-aging and beneficial health effects.

Keywords: antioxidants, Gaddi goats, hill cattle, milk proteins, skim milk, sulfate-polyacrylamide gel electrophoresis.

Immune response evaluation in Balb/c mice after crude extract of Anisakis typica sensitization

Research (Published online: 04-10-2019)

3. Immune response evaluation in Balb/c mice after crude extract of Anisakis typica sensitization

Linda Haryadi, Eddy Suprayitno, Aulanni'am Aulanni'am and Anik Martinah Hariati

Veterinary World, 12(10): 1529-1534

ABSTRACT

Background and Aim: Anisakis is a global challenge for a fish product which may lead to a decrease in economic value and consumers' preference. Skipjack (Katsuwonus pelamis) in Kupang, Nusa Tenggara Timur, Indonesia, have important economic value for local fisheries. Anisakis typica is one of the Anisakis species which potent to induce an allergic reaction. However, the study about A. typica involved in the dendritic cells (DCs), T helper 1 (Th1), T helper 2 (Th2), and regulatory T cells (Tregs) is still limited. This study aimed to analyze the dynamic changed of the immune system including DCs, CD4+ T cells, and Tregs after 1 week of A. typica sensitization.

Materials and Methods: Twenty-four male Balb/C mice were randomly divided into four groups (n=6), mice treated with crude A. typica extract (CAE) 50, 75, and 100 mg/kg BW, respectively. CAE was given orally per day for a week. At the end of the experiment, the animals were sacrificed and the spleen was collected. DCs were labeled as CD11c+ interleukin-6+ (IL-6+); CD4+ T cells were distinguished as Th1 (CD4+ interferon-γ+ [IFN-γ+]) and Th2 (CD4+ IL-4+ and CD4+ IL-5+); Tregs were labeled as CD4+CD25+CD62L+. The expression of each cell was determined by flow cytometry.

Results: Our result described that CAE elicits CD11c+ IL-6+, CD4+ IFN-γ+, CD4+ IL-4+, and CD4+ IL-5+ and reduces CD4+CD25+CD62L+ significantly (p<0.05) in dose-dependent manner in mice after A. typica infection.

Conclusion: The Th1/Th2 ratio after A. typica crude extract treatment exhibits a mixed pattern rather than the classical model allergy to food antigens. Our study is expected as a basic understanding of the changes in immune response after A. typica infection.

Keywords: allergy, dendritic cells, inflammation, nematode, regulatory T cells.

Detection of virulence and multidrug resistance operons in Pseudomonas aeruginosa isolated from Egyptian Baladi sheep and goat

Research (Published online: 04-10-2019)

2. Detection of virulence and multidrug resistance operons in Pseudomonas aeruginosa isolated from Egyptian Baladi sheep and goat

A. N. Dapgh, A. S. Hakim, H. A. Abouelhag, A. M. Abdou and E. A. Elgabry

Veterinary World, 12(10): 1524-1528

ABSTRACT

Background: Pseudomonas aeruginosa is a pit of an enormous group of free-living bacteria that are able to live everywhere and suggested to be the causative agent of great scope of acute and chronic animal infections.

Aim: The current study was carried out to illustrate the prevalence of P. aeruginosa in small ruminants and existence of some virulence operons as well as its antimicrobial resistance.

Materials and Methods: A total of 155 samples from sheep and 105 samples from goats (mouth abscesses, fecal swabs, nasal, tracheal swabs, and lung tissue) were collected for bacteriological study, existence of some virulence expression operons with the study of their sensitivity to the antimicrobials using disc diffusion and presence of mexR operon which is responsible for multidrug resistance (MDR).

Results: The bacteriological examination revealed that P. aeruginosa was isolated from nine out of 155 samples from sheep (5.8%) and four isolates out of 105 samples from goat (3.8%). It is found that 12 (92.3%), 10 (76.9 %), and 8 (61.5%) of P. aeruginosa isolates harbored hemolysin phospholipase gene (pclH), gene (exoS), and enterotoxin gene (toxA), respectively. The results of antibiotic sensitivity test showed that all tested isolates were resistant to ampicillin, bacitracin, erythromycin, streptomycin, tetracycline, trimethoprim-sulfamethoxazole, and tobramycin but sensitive to ciprofloxacin and norfloxacin. The MDR (mexR) operon was existed in all isolates.

Conclusion: There is a growing risk for isolation of virulent MDR P. aeruginosa from sheep and goat illness cases, and this should be regarded in the efficient control programs.

Keywords: drug resistance, goat, Pseudomonas aeruginosa, sheep, virulence.

Seroprevalence and factors associated with bovine and swine toxoplasmosis in Bobo-Dioulasso, Burkina Faso

Research (Published online: 02-10-2019)

1. Seroprevalence and factors associated with bovine and swine toxoplasmosis in Bobo-Dioulasso, Burkina Faso

Dieudonne Tialla, Laibane Dieudonne Dahourou and Oubri Bassa Gbati

Veterinary World, 12(10): 1519-1523

Background and Aim: Toxoplasmosis is a worldwide zoonosis with major public health importance. To know more about this condition in Burkina Faso, this study was implemented to determine the seroprevalence of Toxoplasma gondii infection among pigs and cattle in intra-urban and peri-urban area of Bobo-Dioulasso (Burkina Faso).

Materials and Methods: Serum samples were collected from 600 cattle and 600 pigs with 300 samples from each species in intra-urban and peri-urban area of Bobo-Dioulasso. Data about age, sex, and breed of each animal were also noted. Serum samples were analyzed by indirect enzyme-linked immunosorbent assay to look for immunoglobulin G and immunoglobulin M antibodies to T. gondii.

Results: This study revealed a herd prevalence of 92.5% and 75%, respectively for porcine and bovine toxoplasmosis. At the individual level, we found a prevalence of 29% and 49.2% for cattle and pigs, respectively. For each species, we noticed a significant association between age, sex, breed husbandry system, and the presence of anti-T. gondii antibodies. The prevalence was significantly higher in female, intra-urban system, exotic breed, and animal <2 years old (p<0.05).

Conclusion: The results provided evidence for the presence of T. gondii in pigs and cattle farms around Bobo-Dioulasso. Hence, in Bobo-Dioulasso, raw or undercooked meat consumption is a risk for T. gondii infection for human. Knowledge of the prevalence of toxoplasmosis will help to target prevention efforts.

Keywords: Burkina Faso, cattle, public health, seroepidemiologic studies, swine, toxoplasmosis.