β-lactam resistance in bacteria associated with subclinical mastitis in goats in Thika Subcounty, Kenya

Research (Published online: 25-07-2020)

27. β-lactam resistance in bacteria associated with subclinical mastitis in goats in Thika Subcounty, Kenya

Irene Mkavi Okoko, Naomi Maina, Daniel Kiboi and John Kagira

Veterinary World, 13(7): 1448-1456

ABSTRACT

Aim: This study determined the resistance pattern to β-lactam antibiotics of bacteria isolated from goats with subclinical mastitis in Thika subcounty, Kenya. We also administered a questionnaire to assess the risk factors associated with the occurrence of resistance to commonly used antibiotics.

Materials and Methods: We collected milk samples from 110 lactating dairy goats in Thika subcounty to screen for subclinical mastitis using the California mastitis test. Bacterial isolation and identification were performed according to colony morphology, the hemolytic pattern on sheep blood agar, lactose fermentation on MacConkey plates, Gram staining, and standard biochemical tests. The antibiotic susceptibility of the isolates was determined by the agar disk diffusion method using penicillin G, cephalexin, cefoxitin, and cefotaxime antibiotic disks. The double-disk synergy test using amoxicillin-clavulanic acid was employed as a confirmatory test for extended-spectrum β-lactamase (ESBL) production. Fisher's exact test was used to determine the risk factors associated with the occurrence of antibiotic resistance (p≤0.05 was considered significant).

Results: Of the 110 dairy goats sampled, 72.7% (80) were positive for subclinical mastitis. Isolation and identification of the bacteria from the positive samples yielded 149 bacteria isolates, including Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter spp., Yersinia spp., coagulase-negative staphylococci, and Escherichia coli. A high percentage (76.5%, 114/149) of the bacterial isolates was resistant to at least one of the tested antibiotics. At least 56/106 isolates (52.8%) showing cross-resistance to the β-lactam antibiotics were resistant to all four of the tested antibiotics, while only one isolate was resistant to three antibiotics (penicillin G, cephalexin, and cefoxitin). The double-disk synergy test confirmed that none of the isolates possessed ESBLs. Pre- and post-milking practices (p=0.0336) were found to be significantly associated with the occurrence of antibiotic resistance.

Conclusion: A large proportion of the goats in our study cohort were infected with β-lactam-resistant bacteria associated with subclinical mastitis. Because the identified bacteria are of zoonotic importance, further studies should be undertaken to determine the transmission dynamics between humans and livestock and to identify novel intervention strategies.

Keywords: bacteria, dairy goats, Kenya, subclinical mastitis, β-lactam resistance.

Use of recombinant Brucella outer membrane proteins 19, 25, and 31 for serodiagnosis of bovine brucellosis

Research (Published online: 25-07-2020)

26. Use of recombinant Brucella outer membrane proteins 19, 25, and 31 for serodiagnosis of bovine brucellosis

Aitbay Bulashev, Orken Akibekov, Alfiya Syzdykova, Zhanbolat Suranshiyev and Bakytkali Ingirbay

Veterinary World, 13(7): 1439-1447

ABSTRACT

Background and Aim: Brucellosis remains one of the most common zoonoses. The current anti-brucellosis measures are largely deemed ineffective due to a lack of specificity of conventional serological tests. This study evaluated the use of Brucella outer membrane protein (Omp)19 for serodiagnostic testing.

Materials and Methods: The antigenicity of recombinant Brucella Omp19, Omp25, and Omp31 was examined in serum samples from mice and rabbits immunized with Omp19 or Brucella abortus 19 whole cell (WC) and 12 and 152 cows experimentally or naturally infected with brucellosis, respectively. Serum samples were collected from 151 cows that were vaccinated with B. abortus 19 and 12 unvaccinated heifers that were maintained on a brucellosis-free farm.

Results: Immunization with Omp19 resulted in antibody production in mice after a single injection without the use of adjuvant. Serum antibodies obtained from rabbits immunized with inactivated B. abortus strain 19 WC targeted Omps by enzyme-linked immunosorbent assay (ELISA) and Western blot. Antibodies targeting Omp19 were identified in all B. abortus strain 544 experimentally infected cows at day 14 post-inoculation (p.i.); Omp25 was detected by ELISA at day 28 p.i., while an ELISA targeting Omp31 was negative for 25% of cows at this time point. Omp19 and Omp25 were readily detected by sera from cows from a new epizootic focus. Antibodies recognizing Omps were also detected in >50% of the animals maintained in a brucellosis-free herd at 10 months after vaccination.

Conclusion: Brucella Omp19 in combination with Omp25 and Omp31 may be utilized as target antigens in an ELISA designed for serological testing of unvaccinated cattle.

Keywords: Brucella, diagnosis, enzyme-linked immunosorbent assay, outer membrane proteins.

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.