Antibacterial effects of oak bark (Quercus robur) and heather herb (Calluna vulgaris L.) extracts against the causative bacteria of bovine mastitis

Research (Published online: 28-09-2022)

25. Antibacterial effects of oak bark (Quercus robur) and heather herb (Calluna vulgaris L.) extracts against the causative bacteria of bovine mastitis

Renāte Šukele, Ingus Skadiņš, Rudīte Koka, and Dace Bandere

Veterinary World, 15(9): 2315-2322

ABSTRACT

Background and Aim: Bovine mastitis has a negative impact on animals, and improper antibiotic use has caused an increase in bacterial resistance. Therefore, medicinal plants could serve as an alternative treatment for this condition. Polyphenols have potential as antibiotic agents. Oak bark has long been used as a medicine and has shown antibacterial effects. Moreover, research on heather plant demonstrated that it has antibacterial properties. This study aimed to assess the antibacterial effects of oak (Quercus robur) bark and heather (Calluna vulgaris L.) herb extracts against common bovine mastitis pathogens.

Materials and Methods: Dried oak bark and heather herb were used to prepare extracts using 30%, 50%, and 70% ethanol and acetone as solvents. Their polyphenol content was determined using the Folin–Ciocalteu method. Bovine mastitis-inducing clinical isolates of Escherichia coli, Streptococcus agalactiae, Streptococcus uberis, Serratia liquefaciens, Staphylococcus aureus, and reference cultures of S. aureus and E. coli were used for antibacterial tests. All extracts were screened through a disk diffusion test to ascertain their antibacterial effects, and the minimum inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) were determined for the most effective extracts.

Results: Oak bark extracts had variable antibacterial effects against S. aureus and Streptococcus strains, but no statistically significant difference was observed in activity against E. coli. The disk diffusion test showed that the oak bark extracts obtained using acetone and ethanol at 30% yielded the best results. However, the 70% acetone oak extract alone affected all types of bacteria. Further antibacterial tests of 70% acetone and 30% ethanol oak extracts revealed that the lowest MIC and MBC scores were against S. aureus strains and E. coli reference cultures. Conversely, the heather herb extracts exhibited broader activity against all types of bacteria, although better results were observed against Gram-positive bacteria. There was also a negative correlation between solvent concentration and antibacterial effect (p < 0.05, r = –0.507). The highest inhibition zone scores and broadest spectrum were observed in samples prepared in 30% ethanol. There was no statistically significant correlation between the phenolic content of plants and their antibacterial effects.

Conclusion: Oak bark and heather extracts could be used as potential antibacterial agents against bovine mastitis pathogens.

Keywords: antibacterial, bovine mastitis, Escherichia coli, heather herb, in vitro, oak bark, Staphylococcus aureus, Streptococcus.

Molecular detection and dense granule antigen 6 genotyping of feline Toxoplasma gondii in Phayao, Thailand

Research (Published online: 26-09-2022)

24. Molecular detection and dense granule antigen 6 genotyping of feline Toxoplasma gondii in Phayao, Thailand

Chittakun Suwancharoen, Chorpaka Phuangsri, Khanuengnij Prakhammin, and Ornampai Japa

Veterinary World, 15(9): 2309-2314

ABSTRACT

Background and Aim: Globally, toxoplasmosis is an important zoonotic parasite infection of many warm-blooded animals (including humans). Toxoplasma gondii oocysts are widespread, and their contamination can be primarily attributed to the members of the Felidae family. This study aimed to estimate the prevalence and determine the dense granule antigen 6 (GRA6) genotype of T. gondii among domestic cats in the Phayao Province, Thailand.

Materials and Methods: A total of 124 fecal samples were collected from owned cats in the Muang district, Phayao, Thailand, from January to December 2020. Fecal samples were tested for the presence of T. gondii DNA using targeted B1 gene polymerase chain reaction (PCR) amplification, and positive samples were subsequently analyzed for their T. gondii genotype through PCR-restriction fragment length polymorphism (RFLP) analysis and sequencing of the GRA6 gene.

Results: Among the 124 samples, 46 (37.1%) were tested positive for T. gondii. Only 10 positive DNA samples were successfully amplified for the GRA6 marker. Subsequent PCR-RFLP and sequence analyses indicated that all T. gondii isolates from cats in Phayao belonged to GRA6 genotype I.

Conclusion: Data revealed that toxoplasmosis is remarkably distributed among (studied) domestic cats in Phayao, Thailand. Moreover, the virulent GRA6 allele was found to be circulated among domestic cats in this area. However, no significant correlation was observed between infection rates and different risk factors, which indicated that pet cats of any age, gender, or breed have similar risks of being infected with T. gondii. Our results further suggested that infective oocysts of T. gondii are widely distributed and that environmental contamination with these oocysts will introduce more risks of disease transmission to humans and other animals.

Keywords: domestic cats, genotype, granule antigen 6, Toxoplasma gondii, toxoplasmosis.

Effects of probiotic (Lactobacillus plantarum and Bacillus subtilis) supplementation on mortality, growth performance, and carcass characteristics of native Vietnamese broilers challenged with Salmonella Typhimurium

Research (Published online: 26-09-2022)

23. Effects of probiotic (Lactobacillus plantarum and Bacillus subtilis) supplementation on mortality, growth performance, and carcass characteristics of native Vietnamese broilers challenged with Salmonella Typhimurium

Tran Van Be Nam, Luu Huynh Anh, Huynh Tan Loc, Chau Thi Huyen Trang, Nguyen Thiet, Ly Thi Thu Lan, Tran Hoang Diep, Nguyen Hong Xuan, and Nguyen Trong Ngu

Veterinary World, 15(9): 2302-2308

ABSTRACT

Background and Aim: Probiotic species have been proven to be beneficial on broiler performance; however, most studies have focused on industrial chickens with fast growth, whereas little information concerning the use of these species on native chickens is available. This study aimed to investigate the effects of probiotics Lactobacillus plantarum (LP) and Bacillus subtilis (BS) on the mortality, growth rate, and carcass characteristics in native Noi chickens challenged with Salmonella Typhimurium.

Materials and Methods: We divided 420 1-day-old Noi chicks into seven different treatment groups (n = 60): negative control (no S. Typhimurium, no probiotics or antibiotics); positive control (PC, S. Typhimurium infection, no probiotics or antibiotics); and S. Typhimurium infection and supplementation with LP, BS, LP + BS, enrofloxacin, and commercial probiotics, respectively. Treatment was for 96 days, and the chicks were orally challenged with S. Typhimurium at 22 days old.

Results: No deaths occurred during the 4 weeks post-infection in the negative control, LP, or LP+BS groups. The PC group had the highest mortality rate (20%). Re-isolation of S. Typhimurium from the liver, spleen, and heart showed reduced bacterial counts at 1 week post-infection in the LP, BS, and LP + BS groups. The lowest body weight gain was observed in the PC group (949 g/bird), and chicks in the LP group gained 1148 g/bird. An improved feed conversion ratio was noted in the groups receiving probiotic supplementation (3.42–3.50 kg feed/kg gain). There was little evidence that probiotics affected carcass percentage and related parameters, such as breast, thigh and drumstick, and wings.

Conclusion: Lactobacillus plantarum or BS dietary supplementation to native Noi broilers resulted in a lower mortality rate and improved body weight gain but did not affect carcass characteristics.

Keywords: Bacillus subtilis, growth, Lactobacillus plantarum, mortality, Noi chicken.

First study on virulence genes, antimicrobial resistance, and integrons in Escherichia coli isolated from cage, free-range, and organic commercial eggs in Phayao Province, Thailand

Research (Published online: 25-09-2022)

22. First study on virulence genes, antimicrobial resistance, and integrons in Escherichia coli isolated from cage, free-range, and organic commercial eggs in Phayao Province, Thailand

Achiraya Siriphap, Chittakun Suwancharoen, Watchara Laenoi, Parinya Kraivuttinun, Orasa Suthienkul, and Watsawan Prapasawat

Veterinary World, 15(9): 2293-2301

ABSTRACT

Background and Aim: Antimicrobial resistance (AMR) is a global problem that affects human and animal health, and eggs can act as a vehicle for pathogenic and non-pathogenic resistant bacteria in the food chain. Escherichia coli is an indicator of food contamination with fecal materials as well as the occurrence and levels of AMR. This study aimed to investigate the presence of AMR, integrons, and virulence genes in E. coli isolated from eggshell samples of three egg production systems, from supermarkets in Thailand.

Materials and Methods: A total of 750 hen's egg samples were purchased from supermarkets in Phayao Province: Cage eggs (250), free-range eggs (250), and organic eggs (250). Each sample was soaked in buffered peptone water (BPW), and the BPW samples were incubated at 37°C for 18–24 h. All samples were tested for E. coli by the standard conventional culture method. Then, all identified E. coli were tested for antimicrobial susceptibility to 15 antimicrobial agents by the agar disk diffusion method. All E. coli strains were subsequently found to have virulence genes and Classes 1 and 2 integrons by polymerase chain reaction.

Results: Among the eggshell samples, 91 samples were identified as having E. coli (cage eggs, 24 strains; free-range eggs, 27 strains; and organic eggs, 40 strains). Then, among the E. coli strains, 47 (51.6%) were positive for at least one virulence gene. The proportion of AMR in the eggshell samples was 91.2% (83/91), and streptomycin (STR), ampicillin (AMP), and tetracycline (TET) had a high degree of resistance. Among the E. coli strains, 27 (29.7%) strains were positive for class 1 or 2 integrons, and integron-positive strains were commonly found in STR-, AMP-, and TET-resistant strains. Multidrug resistance (MDR) was detected in 57.1% (52/91) of the E. coli strains, with STR-AMP-TET (5.5%) as the most frequent pattern. The proportion of MDR in cage eggs was 75.0% (18/24), which was higher than in both free-range and organic eggs. On the other hand, 53.2% (25/47) of E. coli carrying virulence genes had MDR, distributed across the production systems as follows: Cage eggs, 76.9% (10/13); free-range eggs, 63.6% (7/11); and organic eggs, 34.8% (8/23).

Conclusion: Escherichia coli was detected in eggshell samples from all three egg production systems. The high level of virulence genes, AMR, and integrons indicated the possibility of dissemination of AMR among pathogenic and commensal E. coli through eggshells. These findings could be a major concern to farmers, food handlers, and consumers, especially regarding raw egg consumption.

Keywords: antimicrobial resistance, eggshells, Escherichia coli, integrons, virulence.

Hematological, biochemical, and serological parameters of experimentally infected rabbits with Trichinella nativa and Trichinella spiralis for early identification of trichinellosis

Research (Published online: 24-09-2022)

21. Hematological, biochemical, and serological parameters of experimentally infected rabbits with Trichinella nativa and Trichinella spiralis for early identification of trichinellosis

Orken S. Akibekov, Alfiya S. Syzdykova, Lyudmila A. Lider, Aibek Kh. Zhumalin, Zhasulan K. Baibolin, Fariza S. Zhagipar, Zhannara Zh. Akanova, Ainur A. Ibzhanova, and Aissarat M. Gajimuradova

Veterinary World, 15(9): 2285-2292

ABSTRACT

Background and Aim: Trichinellosis remains a dangerous disease for humans and animals, which can lead to a lethal outcome. The study of specific body reactions in response to invasion by different types of Trichinella can help in the early diagnosis of the disease. This study aimed to investigate the hematological, biochemical, and serological characteristics of rabbits experimentally infected with trichinellosis, as well as the possibility of using changes in these parameters at various disease stages for early hematological, biochemical, and serological diagnosis of trichinellosis.

Materials and Methods: Three groups of rabbits were orally infected with Trichinella nativa and Trichinella spiralis derived from encysted T. spirtalis larvae in pork muscle samples. The first and second groups were infected with T. nativa and T. spiralis, respectively, while the third group served as control by receiving a physiological solution. An ADVIA 2120i automatic hematology analyzer with a blood smear staining module was used to determine the hematological parameters of rabbits. Antigens were used in an enzyme-linked immunosorbent assay (ELISA) to detect antibodies in the sera of infected rabbits that were supernatants containing excretory-secretory antigens (ES-Ag) and somatic antigen (S-Ag).

Results: The detection of biochemical responses to the invasion of T. nativa and T. spiralis isolates was detected and hematological parameters were featured in two cases. Trichinella nativa increased the number of erythrocytes, neutrophils, eosinophils, monocytes, basophils, and thrombocytes on day 7 in rabbits. Creatine kinase (CK) is regarded as the most important indicator for the early detection of parasite invasion. Blood biochemistry showed no active response to T. spiralis infection. However, counts of erythrocytes, neutrophils, lymphocytes, and CK rose significantly. In both color indicators, the number of thrombocytes decreased. Enzyme-linked immunosorbent assay with ES-Ag and S-Ag of these isolates demonstrated the ability to detect antibodies as early as 7 days after infection, with a significant increase in the marker up to 70 days.

Conclusion: On the 7th day after infection, blood tests of infected animals revealed CK-N-acetyl-cysteine (18.2%) and neutrophils (43%) when infected with T. nativa and neutrophils (26.7%) and lymphocytes (20%) when infected with T. spiralis. These indicators may serve as specific parameters for the early detection of Trichinella spp. invasion.

Keywords: diagnostics, experimental infection, rabbits, Trichinella nativa, Trichinella spiralis, trichinellosis.

First isolation of verocytotoxin-producing Escherichia coli O157:H7 from sports animals in Southern Thailand

Research (Published online: 23-09-2022)

20. First isolation of verocytotoxin-producing Escherichia coli O157:H7 from sports animals in Southern Thailand

Jirarat Songsri, Wanida Mala, Sueptrakool Wisessombat, Kesinee Siritham, Sahida Cheha, Nattita Noisa, Tuempong Wongtawan, and Wiyada Kwanhian Klangbud

Veterinary World, 15(9): 2275-2284

ABSTRACT

Background and Aim: Escherichia coli O157:H7 is enterohemorrhagic E. coli, which produces verocytotoxin or Shiga toxin. It is a well-known cause of severe diseases in humans worldwide. Cattle and other ruminants are the main reservoirs of this organism. Sports animals, such as fighting bulls, riding horses, and fighting cocks, are economic animals in Southern Thailand. This study aimed to identify E. coli O157:H7 from the rectal swabs of these sports animals and determine the antimicrobial susceptibility patterns of isolated bacteria.

Materials and Methods: The rectal swabs were collected from 34 fighting bulls, 32 riding horses, and 31 fighting cocks. The swabs were cultured on MacConkey (MAC) Agar; the suspected colonies were then identified by VITEK® 2 GN card, and the antimicrobial susceptibility was tested by VITEK® 2 AST N194 in VITEK® 2 Compact automation. Escherichia coli O157:H7 was confirmed by culturing on sorbitol MAC agar, the ability to grow at 44°C, and the presence of H7 antigen. In addition, the eaeA (E. coli attaching and effacing), along with stx1 and stx2 (Shiga cytotoxins) genes, were determined using polymerase chain reaction. Finally, the cytotoxicity of Shiga toxin was confirmed using the Vero cytotoxicity test.

Results: Fifty-five suspected isolates (56.70%), which were collected from 19 fighting bulls (55.88%), 13 riding horses (40.63%), and 23 fighting cocks (71.13%), were identified as E. coli. However, one sample (Bull H9/1) from fighting bulls had an equal confidence level (50%) for E. coli and E. coli O157. The confirmation of this isolate demonstrated that it was sorbitol non-fermenter, could assimilate L-lactate, was unable to grow well at 44°C, and reacted with anti-serum to H7 antigen. In addition, it was positive with stx2 and eaeA genes, and the toxin affected Vero cells by a dose-dependent response. The antimicrobial susceptibility test revealed that five out of 55 (9.09%) E. coli isolates were resistant to antimicrobial agents. All five isolates (21.74%) were collected from fighting cocks. Escherichia coli Cock H4/3 was only one of the five isolates resistant to three antimicrobial agents (ciprofloxacin, moxifloxacin, and trimethoprim/sulfamethoxazole). Fortunately, it was not multidrug-resistant bacteria.

Conclusion: This is the first report on detection of E. coli O157:H7 in fighting bulls and antibiotic-resistant characteristic of E. coli in fighting cocks in Southern Thailand. This research is beneficial in preventing the dissemination of E. coli O157:H7 or antimicrobial agent-resistant E. coli in sports animals and humans.

Keywords: Escherichia coli O157:H7, fighting bulls, fighting cocks, riding horses, sport animal, verocytotoxin.

First molecular characterization of capsule expression and antibiotic susceptibility profile of Staphylococcus aureus isolates from bovine mastitis in Jordan

Research (Published online: 23-09-2022)

19. First molecular characterization of capsule expression and antibiotic susceptibility profile of Staphylococcus aureus isolates from bovine mastitis in Jordan

Mohammad Hamdi Gharaibeh and Luay F. Abu-Qatouseh

Veterinary World, 15(9): 2269-2274

ABSTRACT

Background and Aim: Bovine mastitis has long been considered the most important cause of economic losses in the dairy industry. Staphylococcus aureus is the most frequently isolated pathogen from bovine mastitis cases worldwide. Capsular polysaccharides (CPs) of serotype 5 (CP5) or serotype 8 (CP8) are the most prevalent capsule genotypes related to infections associated with S. aureus in humans. However, a variety of CPs has been reported in ruminants and other hosts. Information regarding the relationship between genotypic and phenotypic capsule variation and bovine mastitis in Jordan is scarce. Thus, we aimed to determine the prevalence of S. aureus capsule genotypes CP5 and CP8 in milk from bovine mastitis cases and the antimicrobial susceptibility profile of the recovered isolates in 27 dairy farms in Jordan.

Materials and Methods: Staphylococcus aureus strains were isolated from bovine mastitis cases in two districts of Jordan. All S. aureus isolates were initially identified using conventional biochemical and microbiological methods. Subsequently, confirmation of the identity of S. aureus was performed by polymerase chain reaction (PCR) targeting nuc gene. Capsule polysaccharide typing was performed by PCR specific for CP5 and CP8. In addition, we assessed the antibiotic susceptibility profile of S. aureus isolates against commonly used antimicrobials by the disk diffusion method according to Clinical and Laboratory Standards Institute guidelines.

Results: We collected 148 clinical isolates of S. aureus from bovine mastitis cases in the Zarqa (67.6%, n = 100) and Irbid (32.4%, n = 48) districts. Most isolates possessed capsule genotypes (91.3%), predominantly CP8 (88.6%). Only 8.7% of the isolates were nontypeable by PCR. In addition, we found statistically significant differences between the geographical region and the status of methicillin-resistant capsule genotypes (p < 0.05). The rates of resistance to β-lactam, macrolide, and fluoroquinolone antibiotics were very low, but resistance to tetracyclines was considerably high (22.3%). Significantly, mastitis isolates from Irbid showed a higher rate of resistance to ciprofloxacin (8.3% vs. 0%), while isolates from Zarqa showed a significantly higher rate of resistance to gentamicin (12.0% vs. 6.2%).

Conclusion: We established associations between capsule genotypes and antimicrobial resistance and the pathogenic behavior of S. aureus isolated from bovine mastitis cases. Further studies are necessary to fully elucidate the role and mechanisms of capsular expression in the epidemiological and molecular variability of S. aureus in bovine mastitis.

Keywords: antibiotic susceptibility, bovine mastitis, capsule genotypes, polymerase chain reaction, Staphylococcus aureus.