Prevalence, molecular detection, and virulence gene profiles of Campylobacter species in humans and foods of animal origin

Research (Published online: 24-07-2020)

25. Prevalence, molecular detection, and virulence gene profiles of Campylobacter species in humans and foods of animal origin

Ashraf M. A. Barakat, Khaled A. Abd El-Razik, Hassan A. Elfadaly, Nagwa S. Rabie, Sabry A. S. Sadek and Abdulaziz M. Almuzaini

Veterinary World, 13(7): 1430-1438

ABSTRACT

Background and Aim: Campylobacteriosis is one of the most well-characterized bacterial foodborne infections worldwide that arise chiefly due to the consumption of foods of animal origin such as poultry, milk, and their products. The disease is caused by numerous species within the genus Campylobacter, but Campylobacter jejuni is the most commonly isolated species from established cases of human campylobacteriosis. This study was conducted to determine the prevalence and virulence of Campylobacter isolates from human, chicken, and milk and milk products in Egypt.

Materials and Methods: A total of 1299 samples (547 chicken intestine and liver, 647 milk and milk products, and 105 human stool) were collected and microbiologically investigated, confirmed by multiplex polymerase chain reaction (PCR) targeting the 23S rRNA, hipO, and glyA genes specific for Campylobacter spp., C. jejuni, and Campylobacter Coli, respectively, followed by virulence genes (Campylobacter adhesion to fibronectin F [cadF] and cdtB) detection using PCR.

Results: About 38.09%, 37.84%, and 8.5% of human stool, chicken, and milk and milk product samples, respectively, were bacteriologically positive, with a total of 302 Campylobacter isolates. All isolates were molecularly confirmed as Campylobacter spp. (100%) where 285 isolates (94.37%) were identified as C. jejuni and 17 isolates (5.62%) as C. coli. Regarding the virulence pattern, all isolates (100%) carried cadF gene while cytolethal distending toxin B gene was definite in 284/302 isolates (94%), concisely, 282/285 (98.94%) C. jejuni isolates, and in 2/17 (11.76%) C. coli isolates.

Conclusion: The widespread presence of these highly virulent Campylobacter, especially C. jejuni, proofs the urgent need for the implementation of stringent control, public health, and food protection strategies to protect consumers from this zoonotic pathogen. The availability of information about pathogen virulence will enable enhanced local policy drafting by food safety and public health officials.

Keywords: Campylobacter, Egypt, food, human stool, multiplex polymerase chain reaction, virulence genes.

Artemisia vulgaris efficacies against various stages of Aedes aegypti

Research (Published online: 24-07-2020)

24. Artemisia vulgaris efficacies against various stages of Aedes aegypti

Vika Ichsania Ninditya, Endah Purwati, Ajeng Tyas Utami, Aprillyani Sofa Marwaningtyaz, Nadia Khairunnisa Fairuz, Rini Widayanti and Penny Humaidah Hamid

Veterinary World, 13(7): 1423-1429

ABSTRACT

Background and Aim: Aedes aegypti is the vector of dengue fever, dengue hemorrhagic fever, chikungunya, and, most recently, Zika. Dengue fever is one of Indonesia's endemic diseases. The principal tool for preventing dengue is controlling Ae. aegypti by chemical insecticides since vaccine against dengue is still under research. However, Ae. aegypti developed resistance to various chemical insecticides worldwide. Therefore, research on alternate compounds as mosquito insecticides is urgently needed. This study demonstrated the efficacy of Artemisia vulgaris extract as larvicidal, ovicidal, adulticidal, repellency, and oviposition deterrent activity against Ae. aegypti.

Materials and Methods: A. vulgaris was obtained from Temanggung, Indonesia, while the eggs of Ae. aegypti were collected from Yogyakarta, Indonesia, and were hatched in Laboratory of Parasitology, Faculty of Veterinary Medicine, Universitas Gadjah Mada. Larvicidal activity was evaluated according to the WHO protocol; adulticidal activity was performed using the Centers for Disease Control protocol. Oviposition activity was evaluated using ovitraps added with A. vulgaris extract, complete protection time in the repellent assay was defined as the number of minutes elapsed between compound application and the landing of the first mosquito.

Results: A test of the larvicidal activity of A. vulgaris extract returned an LC50 of 65.8 ppm (r2=0.9014) in 1 h and 18.6 ppm (r2=0.575) in 24 h. A. vulgaris was effective as an adulticidal, demonstrating LC50 values of 11.35 mg (r2=0.875) in 90 min, 9.63 mg (r2=0.924) in 105 min, and 6.46 mg (r2=0.925) in 120 min. A. vulgaris at a concentration of 1000 ppm was able to reach 96% of oviposition deterrent effect. The ovicidal assay, a concentration of 1000 ppm resulted in 82.67% of eggs remaining unhatched. An extract concentration of 80 mg/ml achieved 63.3±3.5% biting repellency in adults.

Conclusion: This study gives a clear indication that A. vulgaris extract acts on Ae. aegypti at various developmental stages and is a potential alternative bioinsecticide for controlling this disease vector.

Keywords: Aedes aegypti, Artemisia vulgaris, bioinsecticide.

Penetration depth study of 830 nm low-intensity laser therapy on living dog tissue

Research (Published online: 23-07-2020)

23. Penetration depth study of 830 nm low-intensity laser therapy on living dog tissue

Naruepon Kampa, Supranee Jitpean, Suvalak Seesupa and Somphong Hoisang

Veterinary World, 13(7): 1417-1422

ABSTRACT

Background and Aim: Recent studies have shown that low-intensity laser therapy (LILT) enhances chronic wound healing, reduces pain, reduces inflammation, and improves post-operative rehabilitation. However, clinical outcomes in the veterinary use of LILT vary between different experimental studies. This is explained by improper laser parameter settings and limits of its penetration depth. This study aimed to investigate the penetration depth of 830 nm LILT on living dog tissue in different operating modes. This entailed continuous wave (CW) versus pulse wave (PW) and with contact versus non-contact techniques of the laser probe at different tissue-laser probe distances. The results can be applied for use in clinical practice.

Materials and Methods: Twenty-four dogs that had undergone abdominal surgery were included in this study. The laser parameters were set at 200 mW, fluence of 4 J/cm2 and the laser power output denoted as mean output power (MOP) was measured by a power meter.

Results: The MOP of the 830 nm CW laser was significantly higher than the PW laser (p<0.05). The MOP of the contact technique was significantly greater than that of the non-contact technique in both CW and PW modes (p<0.05). The MOP through the skin tissue was between 16.09 and 18.60 mW (8.05-9.30%) for the contact technique and 8.73 and 19.36 mW (4.37-9.68%) for the non-contact technique. In the muscle-skin layer, the MOP was between 0.50 and 1.56 mW (0.25-0.78%) and the MOP was not detected using the non-contact technique with a 5 cm tissue-laser probe distance.

Conclusion: Our study indicates that 830 nm LILT (with laser parameter setting at 200 mW, fluence of 4 J/cm2 for both contact and non-contact techniques, and tissue-laser probe distance up to 5 cm) was appropriate for treatments within 14 mm of depth. However, the use of 830 nm LILT for an application in which the target tissue is deeper than 14 mm may limit its positive effect.

Keywords: living dog tissue, low-intensity laser therapy, mean output power, penetration depth.

Detection and antibiotic resistance of Mycoplasma gallisepticum and Mycoplasma synoviae among chicken flocks in Egypt

Research (Published online: 23-07-2020)

22. Detection and antibiotic resistance of Mycoplasma gallisepticum and Mycoplasma synoviae among chicken flocks in Egypt

Marwa Emam, Yousreya Mohamed Hashem, Mahmoud El-Hariri and Jakeen El-Jakee

Veterinary World, 13(7): 1410-1416

ABSTRACT

Background and Aim: Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) are the most significant pathogens of avian mycoplasmosis. This study aimed to isolate and identify MG and MS from chickens and detect the various virulence genes in the isolates. Moreover, the efficacies of different antibiotics were tested to identify suitable treatment regimens.

Materials and Methods: We isolated MG and MS from 487 chicken samples of different ages located in different Governorates in Egypt using conventional isolation methods. The isolates were characterized by polymerase chain reaction (PCR) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and then tested for antibiotic sensitivity by the minimum inhibitory concentration (MIC) method.

Results: The prevalence of MG among the isolates was 9.85%, with the highest percentage isolated from air sacs, while the prevalence of MS among the isolates was 1.6%. Moreover, the highest levels of the prevalence of both MG and MS were during the winter and autumn sampling, while the lowest levels were in the summer and spring. Following the 16S rRNA-based detection of Mycoplasma isolates, 14 MG and 5 MS isolates were identified by different PCR-based detection methods for various virulence genes. Nine MG isolates contain the mgc2 gene, six MG isolates contain the gapA gene, and three MS isolates contain the vlhA gene. We validated a duplex PCR method for the simultaneous identification of MG and MS, based on 100% of the MG and MS isolates generating common bands at 55 and 17 kDa, respectively. The MIC method identified tiamulin and spiramycin as the antibiotics of choice for the treatment of MG and MS infections, respectively.

Conclusion: For more precise diagnosis of Mycoplasma infections in chicken flocks, conventional isolation methods must be confirmed by PCR. SDS-PAGE analysis helps in epidemiological studies and vaccine preparation. The MIC method can be used to help develop therapies to control avian mycoplasmosis infections.

Keywords: gapA gene, mgc2 gene, minimum inhibitory concentration, Mycoplasma infection, sodium dodecyl sulfate, vlhA gene.

Cinnamomum burmannii (Nees & T. Nees) Blume and Eleutherine palmifolia (L.) Merr. extract combination ameliorate lipid profile and heart oxidative stress in hyperlipidemic mice

Research (Published online: 22-07-2020)

21. Cinnamomum burmannii (Nees & T. Nees) Blume and Eleutherine palmifolia (L.) Merr. extract combination ameliorate lipid profile and heart oxidative stress in hyperlipidemic mice

Retno Susilowati and Abdul Malik Setiawan

Veterinary World, 13(7): 1404-1409

ABSTRACT

Background and Aim: Hyperlipidemia is an important risk factor for cardiovascular disease. The use of statins has adverse side effects that result in oxidative stress disorders. The objective of this study was to investigate the antihyperlipidemic effect of a combination of Cinnamomum burmannii and Eleutherine palmifolia extract in high-fat diet (HFD)-induced hyperlipidemia mice.

Materials and Methods: Mice were divided into eight groups (n=4): Control group or healthy mice (normal), HFD-induced hyperlipidemic mice without any treatment (CE0), HFD-induced hyperlipidemic mice treated with 3.6 mg/kg body weight (BW) atorvastatin (atorvastatin), and HFD-induced hyperlipidemic mice treated with a combination of C. burmannii and E. palmifolia in the following ratios: 300:0 (C300), 225:75 (C225), 150:150 (CE150), 75:225 (E225), and 0:300 (E300). Mice were fed a HFD for 4 months to induce hyperlipidemia. Total cholesterol, cholesterol oxidase-peroxidase aminophenazone (CHOD-PAP), triglyceride-glycerine, and fat serum were analyzed with colorimetric method. The measurement of superoxide dismutase was done with the xanthine oxidase method and malondialdehyde measurement was done with the thiobarbituric acid method.

Results: Results showed an increase in antihyperlipidemic characteristics as the concentration of E. palmifolia extract (p<0.05) increased. Duncan's multiple range test also showed an increase in anti-stress oxidation as the concentration of C. burmannii extract (p<0.05) increased.

Conclusion: The E225 group showed the most potential as a safe, antihyperlipidemic agent characterized by improvement in lipid profile and antioxidant balance.

Keywords: antihyperlipidemic, Cinnamomum burmannii, Eleutherine palmifolia, lipid profile, malondialdehyde, superoxide dismutase.

Antimicrobial resistance profiles in bacterial species isolated from fecal samples of free-ranging long-tailed macaques (Macaca fascicularis) living in Lopburi Old Town, Thailand

Research (Published online: 22-07-2020)

20. Antimicrobial resistance profiles in bacterial species isolated from fecal samples of free-ranging long-tailed macaques (Macaca fascicularis) living in Lopburi Old Town, Thailand

Duangjai Boonkusol, Suporn Thongyuan, Nantana Jangsuwan and Pornchai Sanyathitiseree

Veterinary World, 13(7): 1397-1403

Background and Aim: At present, increasing in long-tailed macaques (Macaca fascicularis) population in Lopburi old town caused several problems in its community, in particular with sanitation problem. The present study aimed to explore species distribution and antimicrobial resistance patterns in bacteria isolated from feces of the free-ranging long-tailed macaques (Macaca fascicularis) in Lopburi Old Town, Thailand.

Materials and Methods: Fresh fecal samples were collected from October 2018 to July 2019 from seven troops of macaques. Bacterial colonies were identified based on Gram stain and standard biochemical techniques. Sensitivity toward eight different antibiotics, including amoxicillin, amoxicillin-clavulanate, cephalexin, clindamycin, doxycycline, enrofloxacin, erythromycin, and gentamicin, was analyzed using the disk diffusion method.

Results: A total of 1050 fecal samples were collected. Five unique bacterial species were identified, including Escherichia coli, Enterobacter spp., Proteus spp., Salmonella Group B, and Citrobacter spp. in 100%, 25.71%, 18%, 1.71%, and 0.57% of the fecal specimens, respectively. Among 70 distinct isolates of E. coli, 63 (93%) were resistant to multiple drugs, including amoxicillin, cephalexin, clindamycin, and erythromycin; one isolate (6%) was resistant to clindamycin only. Furthermore, 17 isolates (94%) of Salmonella Group B were resistant to both clindamycin and erythromycin. Five of the six Citrobacter spp. isolates (83%) were also multidrug-resistant (to cephalexin, clindamycin, and erythromycin); the one remaining Citrobacter spp. isolate (6%) was resistant to both clindamycin and erythromycin. However, a high percentage of E. coli, Salmonella Group B and Citrobacter spp. remained susceptible to amoxicillin-clavulanate, enrofloxacin, and doxycycline.

Conclusion: Our findings provide the basic information for the selection of empirical therapy and for the evaluation of the scale of antibiotic resistance associated with macaques living in Lopburi Old Town.

Keywords: antibiotic, drug, monkey, resistant, susceptible.

Harnessing the antibacterial activity of Quercus infectoria and Phyllanthus emblica against antibiotic-resistant Salmonella Typhi and Salmonella Enteritidis of poultry origin

Research (Published online: 21-07-2020)

19. Harnessing the antibacterial activity of Quercus infectoria and Phyllanthus emblica against antibiotic-resistant Salmonella Typhi and Salmonella Enteritidis of poultry origin

Amruta Nair, T. Balasaravanan, Sunil Jadhav, Vysakh Mohan and Chethan Kumar

Veterinary World, 13(7): 1388-1396

Background and Aim: In a scenario of the ineffectiveness of the current drugs against antibiotic-resistant pathogens, the herbal extracts can serve as an alternative remedy. This study appraises the antibacterial potency of Quercus infectoria (gall), Phyllanthus emblica (fruit) individually and synergistically against antimicrobial-resistant (AMR) Salmonella Typhi and Salmonella Enteritidis in a time and dose-dependent manner. Further, the antibacterial phytocompounds were identified employing gas chromatography-mass spectrometry (GC-MS).

Materials and Methods: Preliminary antibacterial activity of the plant extracts was assessed using the agar disk diffusion method. In vitro evaluations of Q. infectoria methanolic extract (QIME) and P. emblica methanolic extract (PEME) against S. Typhi and S. Enteritidis were carried out using plate count method.

Results: QIME and PEME at a dose rate of 50 mg/ml and 25 mg/ml, respectively, had a complete bactericidal effect on AMR S. Typhi and S. Enteritidis whereas 10 log10 CFU/ml of exponential growth was seen in untreated control groups. At the lower concentrations, QIME and PEME had a significant bacteriostatic effect (3-6 log10 reduction of the test isolates). The synergistic antibacterial effect obtained from the combination of these two plant extracts at 12.5 mg/ml was superior (p<0.001) than the individual treatments. Phytochemical profiling indicated the presence of tannins, flavonoids, saponins, and terpenoids in both the plant extracts. GC-MS analysis of QIME and PEME revealed the presence of 16 and 15 antibacterial phytocompounds, respectively. Further 1, 2, 3 Benzenetriol was found as the prominent active principle.

Conclusion: The findings validate that QIME and PEME are potential antibacterial agents against AMR S. Typhi, S. Enteritidis and can play a promising role in antimicrobial packaging, poultry feed additives and can also serve as a platform for formulating effective phytotherapeutics.

Keywords: antimicrobial-resistant, Phyllanthus emblica, phytochemicals, gas chromatography-mass spectrometry, Quercus infectoria, Salmonella.