Comparison of conventional polymerase chain reaction and routine blood smear for the detection of Babesia canis, Hepatozoon canis, Ehrlichia canis, and Anaplasma platys in Buriram Province, Thailand

Research (Published online: 24-05-2019)

12. Comparison of conventional polymerase chain reaction and routine blood smear for the detection of Babesia canis, Hepatozoon canis, Ehrlichia canis, and Anaplasma platys in Buriram Province, Thailand

Rucksak Rucksaken, Cherdsak Maneeruttanarungroj, Thanaporn Maswanna, Metita Sussadee and Pithai Kanbutra

Veterinary World, 12(5): 700-705

ABSTRACT

Background and Aim: Dog blood parasites are important tick-borne diseases causing morbidity and mortality in dogs worldwide. Four dog blood parasites species are commonly found in Thailand: Babesia canis, Hepatozoon canis, Ehrlichia canis, and Anaplasma platys. They are transmitted easily by tick species. However, there is little prevalence data available in Thailand. Diseases presentation of blood parasites infection is similar, but the treatment of each species is different. Current diagnosis mainly relies on microscopic examination of a stained blood smear, which has low sensitivity. Therefore, accurate diagnosis is important. This study aims to evaluate the efficacy of the conventional polymerase chain reaction (PCR) method and routine blood smears in the detection of four blood parasites species in dogs from Buriram Province, Thailand.

Materials and Methods: In total, 49 EDTA-blood samples were collected from dogs in Buriram Province, Thailand. Blood parasite infection was compared using the Giemsa-stained blood smear technique to identify the parasite under a 100× oil immersion with PCR amplification of the 18S rDNA gene of B. canis and H. canis and the 16S rDNA gene of E. canis and A. platys.

Results: Only one dog out of 49 was positive for H. canis based on microscopic examination whereas the PCR results showed that 2.04% (1/49), 4.08% (2/49), 36.73% (18/49), and 30.61% (15/49) of dogs were positive for B. canis, H. canis, E. canis, and A. platys, respectively. Moreover, coinfection was found in 16.33% (8/49) of dogs.

Conclusion: This study is the first report to demonstrate the molecular prevalence of blood parasites in domestic dogs in Buriram Province. The results indicated that the PCR method exhibited much higher sensitivity and reliability for blood parasites diagnosis in dogs. Therefore, our data support serious concern regarding the diagnostic technique used in routine blood testing and also provide prevalence data for the management and control of blood parasites in this area.

Keywords: Blood parasites, dog, polymerase chain reaction, prevalence.

Detection of serum antibodies against Leptospira spp. in brown rats (Rattus norvegicus) from Grenada, West Indies

Research (Published online: 23-05-2019)

11. Detection of serum antibodies against Leptospira spp. in brown rats (Rattus norvegicus) from Grenada, West Indies

Ravindra Nath Sharma, Katelyn Thille, Brianna Piechowski and Keshaw Tiwari

Veterinary World, 12(5): 696-699

ABSTRACT

Background and Aim: Leptospirosis is an emerging disease of animals and humans. Among rodents brown rats (Rattus norvegicus) are an important reservoir of bacteria Leptospira. There is a paucity of information on reservoirs of Leptospira in Grenada. This study was conducted to estimate the prevalence of antibodies against Leptospira spp. in brown rats in a densely human populated area of Grenada.

Materials and Methods: Blood samples from 169 brown rats were collected and sera screened for antibodies against Leptospira spp. using enzyme-linked immunosorbent assay.

Results: Among a total of 169 brown rats trapped in two parishes in Grenada, 77/169 (45.5%) were positive for Leptospira spp. antibodies. A significant difference in seropositive population of brown rats between two collection sites was observed. No differences were found between sex and age of seropositive rats.

Conclusion: Due to the close contact of brown rats with humans in Grenada, rats should be considered a high-risk factor in transmission of Leptospira to humans. Appropriate preventive measures should be instituted to prevent the transmission of Leptospira infection to humans.

Keywords: brown rats, enzyme-linked immunosorbent assay, Grenada, Leptospira spp.

Detection and characterization of clustered regularly interspaced short palindromic repeat-associated endoribonuclease gene variants in Vibrio parahaemolyticus isolated from seafoods and environment

Research (Published online: 21-05-2019)

10. Detection and characterization of clustered regularly interspaced short palindromic repeat-associated endoribonuclease gene variants in Vibrio parahaemolyticus isolated from seafoods and environment

Pallavi Baliga, Malathi Shekar and Moleyur Nagarajappa Venugopal

Veterinary World, 12(5): 689-695

ABSTRACT

Aim: In Vibrio parahaemolyticus, the clustered regularly interspaced short palindromic repeat (CRISPR)-associated cas6 endoribonuclease gene has been shown to exhibit sequence diversity and has been subtyped into four major types based on its length and composition. In this study, we aimed to detect and characterize the cas6 gene variants prevalent among V. parahaemolyticus strains isolated from seafoods and environment.

Materials and Methods: Novel primers were designed for each of the cas6 subtypes to validate their identification in V. parahaemolyticus by polymerase chain reaction (PCR). In total, 38 V. parahaemolyticus strains isolated from seafoods and environment were screened for the presence of cas6 gene. Few representative PCR products were sequenced, and their phylogenetic relationship was established to available cas6 gene sequences in GenBank database.

Results: Of the 38 V. parahaemolyticus isolates screened, only about 40% of strains harbored the cas6 endoribonuclease gene, among which 31.6% and 7.9% of the isolates were positive for the presence of the cas6-a and cas6-d subtypes of the gene, respectively. The subtypes cas6-b and cas6-c were absent in strains studied. Sequence and phylogenetic analysis also established the cas6 sequences in this study to match GenBank sequences for cas6-a and cas6-d subtypes.

Conclusion: In V. parahaemolyticus, the Cas6 endoribonuclease is an associated protein of the CRISPR-cas system. CRISPR-positive strains exhibited genotypic variation for this gene. Primers designed in this study would aid in identifying the cas6 genotype and understanding the role of these genotypes in the CRISPR-cas immune system of the pathogen.

Keywords: cas6 gene, clustered regularly interspaced short palindromic repeats-cas operon, endoribonuclease, type IF system, Vibrio parahaemolyticus.

Golden sea cucumber extract revives glucose transporter-4 and interleukin-6 protein level in diabetic mouse muscle

Research (Published online: 18-05-2019)

9. Golden sea cucumber extract revives glucose transporter-4 and interleukin-6 protein level in diabetic mouse muscle

Bambang Purwanto, Sundari Indah Wiyasihati, Putri Ayu Masyitha, Kristanti Wanito Wigati and Irfiansyah Irwadi

Veterinary World, 12(5): 684-688

ABSTRACT

Background: Streptozotocin (STZ)-induced free radical oxidant activity resulted in muscle wasting due to protein carbonyl (PC), glucose transporter-4 (Glut-4), and interleukin-6 (IL-6) protein alteration. Antioxidant ingredient in the golden sea cucumber extract was found in promising level to inhibit free radical activity.

Aim: This study was aimed to investigate the effect of golden sea cucumber extract on PC, IL-6, and Glut-4 level of STZ-induced diabetes mouse.

Materials and Methods: This study was performed using mice, which were grouped into non-diabetes, diabetes, and diabetes-treated extract groups. The golden sea cucumber was extracted using 70% ethanol, which was administered by oral gavage twice a day for 5 consecutive days.

Results: The extract reduced PC level and improved muscle Glut-4 and IL-6 protein level of diabetic mouse.

Conclusion: The extract of golden sea cucumber revived muscle Glut-4 and IL-6 protein level in protection against muscle wasting.

Keywords: diabetes, glucose transporter, interleukin, muscle.

Prevalence, molecular typing, and antimicrobial resistance of bacterial pathogens isolated from ducks

Research (Published online: 18-05-2019)

8. Prevalence, molecular typing, and antimicrobial resistance of bacterial pathogens isolated from ducks

Hamza M. Eid, Abdelazeem M. Algammal, Wael K. Elfeil, Fatma M. Youssef, Sawsan M. Harb and Ehab M. Abd-Allah

Veterinary World, 12(5): 677-683

ABSTRACT

Aim: This study aimed to investigate the prevalence of different bacterial species affecting ducks as well as demonstrating the antimicrobial susceptibility and molecular typing of the isolated strains.

Materials and Methods: A total of 500 samples were randomly collected from different duck farms at Ismailia Governorate, Egypt. The collected samples were subjected to the bacteriological examination. Polymerase chain reaction (PCR) was applied for amplification of Kmt1 gene of Pasteurella multocida and X region of protein-A (spA) gene of the isolated Staphylococcus aureus strains to ensure their virulence. The antibiotic sensitivity test was carried out.

Results: The most common pathogens isolated from apparently healthy and diseased ducks were P. multocida (10.4% and 25.2%), Escherichia coli (3.6% and 22.8%), Staphylococcus epidermidis (10% and 8.8%), Pseudomonas aeruginosa (2% and 10%), and Proteus vulgaris (0.8% and 10%), respectively. In addition, S. aureus and Salmonella spp. were isolated only from the diseased ducks with prevalence (12.2%) and (2.8%), respectively. Serotyping of the isolated E. coli strains revealed that 25 E. coli strains were belonged to five different serovars O1, O18, O111, O78, and O26, whereas three strains were untypable. Salmonella serotyping showed that all the isolated strains were Salmonella Typhimurium. PCR revealed that four tested P. multocida strains were positive for Kmt1 gene with specific amplicon size 460 bp, while three strains were negative. In addition, all the tested S. aureus strains were positive for spA gene with specific amplicon size 226 bp. The antibiotic sensitivity test revealed that most of the isolated strains were sensitive to enrofloxacin, norfloxacin, and ciprofloxacin.

Conclusion: P. multocida is the most predominant microorganism isolated from apparently healthy and diseased ducks followed by E. coli and Staphylococci. The combination of both phenotypic and genotypic characterization is more reliable an epidemiological tool for identification of bacterial pathogens affecting ducks.

Keywords: Antibiotic sensitivity, duck, Escherichia coli, Pasteurella multocida, polymerase chain reaction, Staphylococci.

Phosphatidylcholine from krill increases plasma choline and its metabolites in dogs

Research (Published online: 16-05-2019)

7. Phosphatidylcholine from krill increases plasma choline and its metabolites in dogs

Lena Burri, Knut Heggen and Andreas Berg Storsve

Veterinary World, 12(5): 671-676

Background and Aim: Choline and its metabolites have multiple physiological roles in the body, which are important for muscle function, memory, methylation reactions, and hepatic lipid transport. This study aimed to investigate, if inclusion of phosphatidylcholine (PC) from Antarctic krill (Euphausia superba) can increase the concentration of choline and its metabolites in plasma of sled dogs in comparison to a control group.

Materials and Methods: Ten adult Alaskan Huskies of both genders were supplemented with PC from 8% dietary krill meal inclusion for 6 weeks, while another ten dogs received no krill meal supplementation. Blood measurements of the two groups were taken at baseline and end of the study and compared for choline and its metabolite concentrations.

Results: The choline concentration of the krill meal-supplemented dogs was significantly higher after 6 weeks of krill meal feeding compared to the control group (mean increase = 4.53 μmol/L in the supplemented versus 1.21 μmol/L in the control group, p=0.014). Furthermore, krill meal-supplemented dogs showed significantly more pronounced increases in betaine (p<0.001), dimethylglycine (p<0.01), trimethylamine-N-oxide (p<0.001), and trimethyllysine (p<0.001) compared to the control group. Significant correlations between changes in choline and changes in its metabolites were observed.

Conclusion: The results showed that krill meal supplementation was associated with significantly higher plasma choline concentrations, which correlated with changed concentrations of choline metabolites.

Keywords: choline metabolites, choline, dog, krill meal, phosphatidylcholine.

Vibriosis and Aeromonas infection in shrimp: Isolation, sequencing, and control

Research (Published online: 14-05-2019)

6. Vibriosis and Aeromonas infection in shrimp: Isolation, sequencing, and control

Hanaa Mohamed Fadel and Maather El-Lamie M.M.

International Journal of One Health, 5: 38-48

Background and Aim: Shrimp is one of the most commonly consumed types of seafood. It is a very nutritious healthy food. Shrimp is low in calories and rich in protein and healthy fats. It also contains a treasure trove of vitamins and minerals. On the negative side, it may be affected by many bacterial diseases which affect its health. Furthermore, it may be incriminated as a vector of foodborne illnesses that range from mild gastrointestinal upset to life-threatening diseases. This study was designed to assess the clinical picture and zoonotic importance of vibriosis and Aeromonas infection in live shrimp and to study the antibacterial effect of citric acid (lemon juice) and acetic acid (vinegar) on these pathogens.

Materials and Methods: A total of 170 live shrimp (Metapenaeus monoceros) samples were collected from Suez City, Egypt. The samples were examined clinically, and then, they were enriched into alkaline peptone water and cultivated on thiosulfate-citrate-bile salts-sucrose agar and ampicillin MacConkey agar for the isolation of Vibrio and Aeromonas species, respectively. The recovered isolates were confirmed biochemically and genotypically using duplex polymerase chain reaction (PCR) and sequencing. The germicidal effects of vinegar and lemon on artificially contaminated shrimp samples with Aeromonas hydrophila and Vibrio parahaemolyticus at different times (0.25, 1, 1.5, and 24 h) and temperatures (5° and 30°C) were studied.

Results: The results revealed that some of the infected shrimp were hypoxic, lethargic with abnormal swimming behavior. In most cases, body appendages, telsons, uropods, and gills took black coloration. In addition, the hepatopancreas appeared soft, swollen, and congested. The prevalence rates of vibriosis in each of the musculature and hepatopancreas were 4.7%, while the prevalence rates of Aeromonas infection in the musculature and hepatopancreas were 11.8% and 11.2%, respectively. Duplex PCR showed that Aeromonas isolates gave double bands: 237 bp specific for gcat and 500 bp specific for 16S rRNA, while Vibrio spp. and Plesiomonas shigelloides isolates gave single band at 500 bp. The effect of organic acid treatment showed that acetic acid (vinegar 5%) had increasing reduction rates that reached its maximum level after 24 h; where it caused (100% inhibition) for A. hydrophila at both temperatures and (33.63% and 60% inhibition) for V. parahaemolyticus at refrigerator and room temperatures, respectively. Moreover, acetic acid was more effective at room temperature than at refrigerator temperature. Concerning the effect of lemon juice (citric acid), it was more effective than acetic acid at short marination (0.25 and 1 h) at both temperatures for the two pathogens. Moreover, lemon was more effective at refrigerator temperature than at room temperature at the same aforementioned time. The difference between the reduction effects of the two acids on both pathogens was statistically significant (p<0.0001).

Conclusion: Overall, the examined shrimp samples were found to be vectors for Vibrio and Aeromonas spp. Application of hygienic measures during handling and cooking of shrimp should be esteemed. The organic acid treatment trial showed that vinegar and lemon juice can be used as a safe and economic method to limit the microbial contamination in seafood.

Keywords: Shrimp, vibriosis, Aeromonas infection, sequencing, decontamination, citric acid, acetic acid.