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.

Susceptibility of virulent and resistant Escherichia coli strains to non-polar and polar compounds identified in Microplumeria anomala

Research (Published online: 21-07-2020)

18. Susceptibility of virulent and resistant Escherichia coli strains to non-polar and polar compounds identified in Microplumeria anomala

Livia Roberta Piedade Camargo, Vania Maria de Carvalho, Ingrit Elida Collantes Díaz, Mateus Luís Barradas Paciencia, Sergio Alexandre Frana, Riad Naim Younes, Antonio Drauzio Varella, Luiz Fernando Lima Reis and Ivana Barbosa Suffredini

Veterinary World, 13(7): 1376-1387

ABSTRACT

Background and Aim: Escherichia coli is one of the main pathogens responsible for veterinary and human infections, and it is associated with significant economic losses in the livestock, as it causes severe diseases to humans, particularly in children. For that reason, there is a need for introducing new drugs to treat E. coli diseases. The Brazilian species richness is a source of potential new antibacterial natural products. The study aimed at the biological and chemical investigation of the organic extract obtained from the stem of Microplumeria anomala (Apocynaceae), EB127, as it was identified as a potential source of new antibacterial compounds to be used in Veterinary.

Materials and Methods: The antibacterial activity was evaluated by disk diffusion and microdilution assays; chromatography, nuclear magnetic resonance spectrometry, and mass spectrometry were used in the isolation and identification of compounds.

Results: EB127 showed activity against E. coli ATCC25922, and against three E. coli strains that were isolated from frigarte's cloaca, named 31/1A, 35A, and 51A. Lupeol, 3-acetyl-11-oxo-β-amyrin, 3-acetyl-11-oxo-α-amyrin, sitosterol, stigmasterol, 3β,7α-dihydroxy-cholest-5-ene, 3β-hydroxy-cholest-5-en-7-one, and 3β-hydroxy-cholest-5,22-dien-7-one were identified in fraction Hex/CHCl3, while loganin, loganic acid, methylanomaline, and anomaline were all identified in EB127 and protocatechuic acid hexoside, ferulic acid, secoxyloganin, feruloylquinic acid, vanillic acid hexoside, protocatechuic acid-4-O-β-hexoside, and rosmarinic acid were tentatively identified in fraction 10%ACN/H2O. E. coli 51A (virulent/non-resistant) showed sensitivity to the antibacterial action of fraction Hex/CHCl3 which contains alkaloids, triterpenes, and steroids, while E. coli 35A (resistant/non-virulent) were more susceptible to 10%ACN/H2O, which contains iridoids as loganin and loganic acid, and glycosylated and non-glycosylated caffeic acids.

Conclusion: Fraction 10%ACN/H2O is of interest in pursuing new drugs to treat resistant E. coli, in veterinary. All compounds were isolated from the plant for the first time and have shown potential as new antibacterial natural products from Amazon plants to be used in veterinary and human diseases.

Keywords: antibacterial agents, companion animals, livestock, plant extracts, poultry, tropical rainforest.

Isolation of peripheral blood mononuclear cells and the expression of toll-like receptors in Betong chickens

Research (Published online: 18-07-2020)

17. Isolation of peripheral blood mononuclear cells and the expression of toll-like receptors in Betong chickens

Anutian Suklek, Autchara Kayan, Jatuporn Rattanasrisomporn and Chaiwat Boonkaewwan

Veterinary World, 13(7): 1372-1375

ABSTRACT

Background and Aim: Toll-like receptors (TLRs) comprise microbial sensing receptors present on cell surfaces that are capable of detecting pathogens. The present study aims to examine the expression of TLRs within the peripheral blood mononuclear cell (PBMC) of the Betong chickens.

Materials and Methods: Blood samples were harvested from 12 Betong (KU line) chickens. Hematological values were calculated. PBMC was isolated from the blood utilizing a Histopaque solution and stored in a RPMI1640 culture medium. Cell viability was investigated using a Trypan Blue dye exclusion test. DNA was extracted from PBMC and the expression of the DNA's TLRs was examined using a polymerase chain reaction.

Results: Hematological values were determined from the blood samples collected in this study obtained from healthy Betong chickens. PBMC that was isolated from the Betong chickens possessed cell viability higher than 95% (95.37±1.06). From the examination of TLRs gene expression, results revealed instances of TLR1.1, TLR1.2, TLR2.1, TLR2.2, TLR3, TLR4, TLR5, TLR 7, TLR15, and TLR21 that were present in the PBMC of Betong chickens.

Conclusion: PBMC isolated from the blood of healthy Betong chickens possessed excellent cell quality. All chicken TLRs were discovered within the PBMC of Betong chickens. Hence, PBMC stands out as one of the premier sources for in vitro studies of chicken immune response.

Keywords: Betong chicken, peripheral blood mononuclear cell, toll-like receptor.

Microbiological quality of beef, mutton, and water from different abattoirs in the Eastern Cape Province, South Africa

Research (Published online: 18-07-2020)

16. Microbiological quality of beef, mutton, and water from different abattoirs in the Eastern Cape Province, South Africa

Philisani Ncoko, Ishmael Festus Jaja and James Wabwire Oguttu

Veterinary World, 13(7): 1363-1371

ABSTRACT

Background and Aim: Abattoir processes from skinning, evisceration, to chilling usually lead to meat contamination by foodborne pathogens. Hence, continual microbial surveillance of slaughter carcasses by veterinary public health officials is key to preventing contamination and outbreak of meat-related foodborne diseases. This study was conducted to determine the Enterobacteriaceae count and aerobic plate count (APC) and to detect Escherichia coli and Salmonella spp. in meat and water from selected slaughter facilities.

Materials and Methods: Retrospective data (n=100) collected in 2017 by the Provincial Veterinary Department of the Eastern Cape Province from abattoirs and prospective survey data of meat (n=50) collected in 2018 from abattoirs in the Eastern Cape Province were utilized in this study. APC and Enterobacteriaceae were enumerated from the samples. In addition, Salmonella and E. coli were isolated from samples using selective media.

Results: The APC in both retrospective and prospective studies for all samples ranged between 2 and 4.50 log CFU/cm2; similar counts of 2-4.00 log CFU/cm2 were recorded for Enterobacteriaceae. No significant difference (p>0.05) for APC and Enterobacteriaceae count across all meat types was noted. Salmonella and E. coli were detected in 50% of beef. E. coli was not detected from mutton, but Salmonella was found in 66.7%. Moreover, 91.7% of the water samples had E. coli, but none had Salmonella.

Conclusion: The levels of Enterobacteriaceae and APC observed in meat satisfy regulatory conditions outlined by the Department of Agriculture, Forestry and Fisheries, South Africa and show that meat produced from these abattoirs is of acceptable microbial quality. However, the quality of water used in the abattoirs does not meet the requirements set by the government, and contributes to contamination of meat produced in the abattoirs under study. Therefore, we recommend that sources of water be continuously investigated to eliminate or reduce the risk of contamination of meat processed in the abattoirs.

Keywords: contamination, foodborne pathogens, hygiene, meat spoilage, water quality.