Characterization and identification of charcoal of inedible Kerandang fish (Channa pleurophthalmus Blkr) body parts and potential antiallergenic properties

Research (Published online: 30-07-2020)

31. Characterization and identification of charcoal of inedible Kerandang fish (Channa pleurophthalmus Blkr) body parts and potential antiallergenic properties

Aryani Aryani, Eddy Suprayitno, Bambang Budi Sasmito and Hardoko Hardoko

Veterinary World, 13(7): 1480-1486

ABSTRACT

Background and Aim: The study about the antiallergenic properties of inedible fish body parts is still limited. Therefore, this study aimed to characterize the charcoal from the body parts of Kerandang fish (Channa pleurophthalmus Blkr) and identify its antiallergenic properties.

Materials and Methods: This study used some non-edible body parts extracted from the Kerandang fish (i.e., the scalp, scales, and dorsal, pectoral, ventral, anal, and caudal fins) using a maceration method with different solvents (ethanol, ethyl acetate, and chloroform). The identification of active compounds in the extract was carried out using liquid chromatography– high-resolution mass spectrometry (LC-HRMS) analysis, while the antihyaluronidase activity was determined using the antihyaluronidase test. The highest charcoal antihyaluronidase activity-extract was applied to ovalbumin-induced mice for 7 days with various doses (10, 15, and 20 mg/kg). The specific immunoglobulin E (IgE) was measured using enzyme-linked immunosorbent assay on day 8.

Results: Our LC-HRMS analysis showed that the active compound of charcoal in the caudal fins of Kerandang fish was hexadecanamide. The highest inhibition (IC50) of hyaluronidase was found in the ethyl acetate extract of fish caudal fins at a concentration of 4 mg/mL. We found that 15 mg/kg body weight of charcoal of fish caudal fins suppressed IgE expression in male mice.

Conclusion: Our findings indicate that the charcoal of non-edible body parts of Kerandang and one of its constituent, hexadecanamide, may have strong antiallergic effects.

Keywords: anti-allergy, charcoal, hyaluronidase, Kerandang fish.

Emergence of Raoultella ornithinolytica isolated from chicken products in Alexandria, Egypt

Research (Published online: 29-07-2020)

30. Emergence of Raoultella ornithinolytica isolated from chicken products in Alexandria, Egypt

Sara M. El-Shannat, Ashraf A. Abd El-Tawab and Wafaa M. M. Hassan

Veterinary World, 13(7): 1473-1479

ABSTRACT

Background and Aim: Raoultella ornithinolytica is one of the emerging gram-negative bacteria, which associated with foodborne illness. Researches affirmed that distinguish between R. ornithinolytica and Klebsiella oxytoca are difficult, as they are phylogenetic related. The evolution of multidrug resistance of Raoultella strains gained more concern for recognition of the pathogen which supports in controlling the disease and minify its threat. This study sought to find a reliable tool for the identification of Raoultella ornithinolytica, isolated from chicken product samples, and assessed the resistance profile of R. ornithinolytica using antibiogram sensitivity tests.

Materials and Methods: Forty samples of chicken products were collected between January and September 2019 from different markets in Alexandria Governorate, Egypt. The products included nuggets, strips, burgers, luncheon meats, pane, frankfurters, and minced chicken meat. The samples were transferred to the Reference Laboratory. The samples were subjected to isolation, biochemical reaction testing, phenotypic system analytical profile index (API) E20, and a detection of antimicrobial susceptibility test. Phenotypic identification was confirmed through matrix-assisted laser-desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS).

Results: Thirty-three bacterial isolates (82.50%) out of 40 samples were isolated into pure cultures from the chicken samples. Three isolates (9.09%) were positive for R. ornithinolytica, while 30 isolates (90.91%) exhibited growth characters for different pathogens (Escherichia coli, Enterobacter aerogenes, Proteus vulgaris, R. ornithinolytica, and Klebsiella pneumoniae). The isolates of R. ornithinolytica were resistant to five types of antibiotics and sensitive to two types of antibiotics.

Conclusion: This study reported the first case of R. ornithinolytica found in chicken products in Egypt. Phenotypic system API 20E and MALDI-TOF MS were found to be reliable tools for confirming the diagnosis of R. ornithinolytica. As it provides rapid identification with high sensitivity and specificity for R. ornithinolytica, which often do not require a molecular procedure for confirmation.

Keywords: analytical profile index 20E, matrix-assisted laser-desorption ionization time-of-flight mass spectrometry, phenotypic system, Raoultella ornithinolytica.

Low genetic diversity among Francisella-like endosymbionts within different genotypes of Hyalomma dromedarii ticks infesting camels in Saudi Arabia

Research (Published online: 28-07-2020)

29. Low genetic diversity among Francisella-like endosymbionts within different genotypes of Hyalomma dromedarii ticks infesting camels in Saudi Arabia

Haitham Elbir, Faisal Almathen and Ayman Elnahas

Veterinary World, 13(7): 1462-1472

ABSTRACT

Background and Aim: Hyalomma dromedarii ticks are vectors of disease agents and hosts of Francisella-like endosymbionts (FLEs). Knowledge about intraspecific genetic variation among H. dromedarii and its Francisella species is limited. The aims of this study were to investigate whether certain H. dromedarii genotypes are specialized in carrying specific Francisella species genotypes and scrutinize the population structure of H. dromedarii ticks in Saudi Arabia.

Materials and Methods: We collected 151 H. dromedarii ticks from 33 camels from 13 locations in Saudi Arabia. The second internal transcribed spacer (ITS2), cytochrome c oxidase subunit-1(COI), and 16S rRNA genes were used for single-and multi-locus sequence typing and phylogenetic analyses. H. dromedarii-borne Francisella was screened using the tul4 gene and 16S rRNA Francisella-specific primers followed by amplicon Sanger sequencing.

Results: Single-locus typing of ticks using ITS2, 16S rRNA, and COI genes yielded 1, 10, and 31 sequence types (ST), respectively, with pairwise sequence similarity of 100% for ITS2, 99.18-99.86% for COI, and 99.50-99.75% for 16S rRNA. COI sequence analysis indicated a lack of strict geographical structuration, as ST15 was found in both Saudi Arabia and Kenya. In contrast, multilocus sequence typing resolved 148 H. dromedarii ticks into 39 genotypes of ticks and three genotypes of FLEs. The ST2-FLE genotype was carried by the tick genotype ST35, while the ST1-FLE genotype and 41.89% of the ST3-FLE genotype were carried by the tick genotype ST32. Accordingly, there appeared to be no specialization of certain tick genotypes to harbor-specific FLE genotypes.

Conclusion: For the 1st time, we have provided an overview of the population structure of H. dromedarii ticks and FLE strains. We found a low level of genetic diversity among FLEs and non-specialized circulation of FLEs among H. dromedarii ticks.

Keywords: camel, endosymbionts Francisella typing, Hyalomma dromedarii.

Malacca leaf ethanolic extract (Phyllanthus emblica) as a hepatoprotector of the liver of mice (Mus musculus) infected with Plasmodium berghei

Research (Published online: 27-07-2020)

28. Malacca leaf ethanolic extract (Phyllanthus emblica) as a hepatoprotector of the liver of mice (Mus musculus) infected with Plasmodium berghei

Nuzul Asmilia, Dwinna Aliza, Yudha Fahrimal, Mahdi Abrar and Sulaiman Ashary

Veterinary World, 13(7): 1457-1461

ABSTRACT

Background and Aim: Although existing research confirms the antiparasitic effect of the Malacca plant against Plasmodium, its effect on the liver, one of the target organs of Plasmodium has not been investigated. Therefore, this study was conducted to explore the potential of the ethanolic extract of Malacca (Phyllanthus emblica) leaves in preventing liver damage in mice (Mus musculus) caused by Plasmodium berghei infection.

Materials and Methods: This study was conducted using the livers of 18 mice fixed in 10% neutral-buffered formalin. A completely randomized design with a unidirectional pattern comprising six treatments was used in this study, with each treatment consisting of three replications. Treatment 0 was the negative control group infected with P. berghei, treatment 1 was the positive control group infected with P. berghei followed by chloroquine administration at a dose of 5 mg/kg BW, and treatments 2, 3, 4, and 5 were groups infected with P. berghei and administered Malacca leaf ethanolic extracts at doses of 100, 300, 600, and 1200 mg/kg BW, respectively. The extracts were administered orally using a gastric tube for 4 consecutive days. Mice were sacrificed on the 7th day and livers were collected for histopathological examination.

Results: Histopathological examination of the livers of mice infected with P. berghei demonstrated the presence of hemosiderin, hydropic degeneration, fat degeneration, necrosis, and megalocytosis. However, all these histopathological changes were reduced in the livers of P. berghei-infected mice treated with various doses of Malacca leaf ethanolic extract. The differences between the treatments were found be statistically significant (p<0.05).

Conclusion: Ethanolic extract of Malacca leaves has the potential to protect against liver damage in mice infected with P. berghei. The dose of 600 mg/kg BW was found to be the most effective compared with the doses of 100, 300, and 1200 mg/kg BW.

Keywords: hepatoprotector, Malacca leaf extract, megalocytosis, Plasmodium berghei.

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.