Development of multiplex reverse transcription-polymerase chain reaction for differentiation of strains of infectious bursal disease virus and primary screening of the virus in Thailand

Research (Published online: 14-12-2021)

5. Development of multiplex reverse transcription-polymerase chain reaction for differentiation of strains of infectious bursal disease virus and primary screening of the virus in Thailand

Nataya Charoenvisal

Veterinary World, 14(12): 3105-3110

ABSTRACT

Background and Aim: A new set of primers (400 base pairs partial of VP2) was designed and used for the infectious bursal disease virus (IBDV) screening test. Using this new primer set, the enzymes MboI and BstNI were unable to differentiate the field and vaccine strains. As a result, a new simple, cheap, and appropriate tool for strain differentiation is required. The objective of this study was to develop the appropriate restriction fragment length polymorphism (RFLP) and multiplex reverse transcription-polymerase chain reaction (RT-PCR) for the differentiation of classic IBDV (cIBDV) strains and very virulent IBDV (vvIBDV) strains in Thailand.

Materials and Methods: Ninety seven bursa of Fabricius from 16 farms were collected from farms in the eastern and central regions of Thailand. RT-PCR screening showed that 82 samples were positive for IBDV and 15 samples were negative. Then, selected samples were sequenced from each farm with a positive test.

Results: The sequencing results showed that samples from six of the farms were vvIBDV and samples from the other six farms were cIBDV. Although the whole genome sequencing was incomplete, both the sequencing results of segment A and segment B showed high similarity between cIBDV and vvIBDV. Restriction enzyme cutting site and primers for multiplex RT-PCR were hard to design. An RT-PCR-RFLP method was developed, but it failed to differentiate IBDV strains. However, the multiplex RT-PCR was able to differentiate cIBDV from vvIBDV. Four primers were used in the multiplex RT-PCR.

Conclusion: These four primers were used together in one reaction at an annealing temperature of 45°C. Therefore, multiplex RT-PCR is a less complicated, cheaper, and less time-consuming method for the differentiation of cIBDV and vvIBDV strains.

Keywords: infectious bursal disease virus, multiplex reverse transcription-polymerase chain reaction, restriction fragment length polymorphism, strain, Thailand.

Antimicrobial resistance profile of extended-spectrum beta-lactamases, adenosine-monophosphate-cyclic, and carbapenemase-producing Gram-negative bacteria isolated from domestic animals

Research (Published online: 13-12-2021)

4. Antimicrobial resistance profile of extended-spectrum beta-lactamases, adenosine-monophosphate-cyclic, and carbapenemase-producing Gram-negative bacteria isolated from domestic animals

Jôiciglecia Pereira dos Santos, Valesca Ferreira Machado de Souza, Marcos Wilker da Conceição Santos, Juliany Nunes dos Santos, Natilene Silva dos Santos, Angélica Prado de Oliveira, Valquíria Tatiele da Silva Rodrigues, Ianei Oliveira Carneiro and Layze Cilmara Alves da Silva Vieira

Veterinary World, 14(12): 3099-3104

ABSTRACT

Background and Aim: The production of beta-lactamase enzymes, such as extended-spectrum beta-lactamase (ESBL), adenosine-monophosphate-cyclic (AmpC), and Klebsiella pneumoniae carbapenemase (KPC), is one of the most important mechanisms of bacterial resistance to antimicrobials. Gram-negative bacteria show significant resistance due to various intrinsic and acquired factors. These intrinsic factors include low permeability of the outer membrane, various efflux systems, and the production of beta-lactamases, while acquired factors include chromosomal mutation and acquisition of resistance genes by horizontal transfer. Mobile elements such as plasmids, integrative conjugative elements, mobilizable islands, or transposable elements are involved in horizontal transfer. At present, the Gram-negative pathogens of most concern are Acinetobacter baumannii, Pseudomonas aeruginosa, and those belonging to the Enterobacteriaceae family (e.g., Escherichia coli, K. pneumoniae, and Proteus mirabilis). This study aimed to evaluate the profile of antimicrobial resistance and the production of the enzymes ESBL, AmpC, and KPC, in 21 gram-negative bacteria isolated from domestic animals treated at the University Veterinary Hospital (HVU) of the Federal University of Western Bahia (UFOB).

Materials and Methods: The biological samples (21) were inoculated to brain heart infusion broth, blood agar, and MacConkey agar and incubated for 24-72 h at 37°C. Gram staining and identification through biochemical tests and matrix-associated laser desorption/ionization time-of-flight mass spectrometry were conducted. To evaluate the antimicrobial resistance profile, the disk diffusion method was used, and 25 antibiotics were employed. For the detection of ESBL, the disk approximation method was applied using chromogenic agar. The presence of KPC was observed using chromogenic agar and the Hodge test. For AmpC evaluation, the disk approximation method was used.

Results: The most isolated agent was E. coli (66.66%, 14/21), followed by K. pneumoniae and P. mirabilis (both 14.29%, 3/21), and then Pasteurella spp. (4.76%, 1/21). The bacterial isolates showed high levels of resistance against clindamycin, penicillin, imipenem, polymyxin, cefoxitin, gentamycin, cefotaxime, ceftazidime, cephalothin, ceftriaxone, ciprofloxacin, trimethoprim/sulfamethoxazole, chloramphenicol, and tetracycline. The best effectiveness rates were observed for cefepime, streptomycin, amoxicillin-clavulanate, aztreonam, nalidixic acid, tobramycin, levofloxacin, amikacin, and meropenem. All biological isolates showed multiple resistance to at least three of the antibiotics tested (3/25), and some showed resistance to 24 of the antibiotics tested (24/25). Among the 21 pathogens analyzed, 8 were ESBL producers (38.09%); of these, 6 were identified as E. coli (28.57%), and 2 were identified as K. pneumoniae (9.52%). Two strains of K. pneumoniae produced both ESBL and KPC. None of the isolates were producers of AmpC.

Conclusion: The results found in the present work raise concern about the level of antimicrobial resistance among pathogens isolated from domestic animals in Brazil. The results highlight the need for the development and implementation of anti-resistance strategies to avoid the dissemination of multiresistant pathogens, including the prudent use of antimicrobials and the implementation of bacterial culture, antimicrobial sensitivity, and phenotypic tests for the detection of beta-lactamase enzymes in bacteria isolated from animals.

Keywords: antimicrobial resistance, extended-spectrum beta-lactamase, Klebsiella pneumoniae carbapenemase, phenotypic tests.

Assessment of the contamination of Lactuca sativa L. (lettuce) and Lycopersicon esculentum (tomato) by pesticides: Case of market gardeners in Ouagadougou

Research (Published online: 11-12-2021)

15. Assessment of the contamination of Lactuca sativa L. (lettuce) and Lycopersicon esculentum (tomato) by pesticides: Case of market gardeners in Ouagadougou

Sibiri Sylvain Rouamba, François Tapsoba, Bazoin Sylvain Raoul Bazié, Ollo Youl, Saybou Savadogo, Elie Kabré, Lassana Sangaré and Aly Savadogo

International Journal of One Health, 7(2): 251-256

ABSTRACT

Background and Aim: The use of pesticides in vegetable production can cause public health problems because these agrochemicals can leave residues in foodstuffs and disrupt the appropriate functioning of the organism. The aim of this study was to assess the level of contamination of lettuce and tomato by pesticide residues.

Materials and Methods: This study was conducted from February 12 to May 3, 2021, in concerned five market gardening sites in the city of Ouagadougou. A survey was conducted among 58 market gardeners selected at random from the sites of Boulmiougou, Tanghin No. 1 and 2, Bissigin, and the National School of Public Health. Simultaneously, 25 lettuce samples and 25 tomato samples were collected in a randomized manner for the examination and determination of pesticide residues using the quick, easy, cheap, effective, rugged, and safe method.

Results: The market gardening population was predominantly represented by women who had a low level of education. The cultivated species were a mixture of exotic cultures and traditional cultures. These crops were treated with various pesticides, the most common of which belonged to the organochlorine family. The tomato was the crop most contaminated by pesticide residues. However, no active ingredient from the synthetic pyrethroid family was found on lettuce or tomato.

Conclusion: Raising the awareness of market gardeners on good practices in the use of pesticides and on alternative methods to synthetic phytosanitary products is of paramount importance for the health of consumers.

Keywords: lettuce, market gardeners, Ouagadougou, pesticides, tomato.

Levels of insulin-like growth factor in bovine, goat, and sheep milk in different lactation periods: The etiological factor of cancer in humans

Research (Published online: 10-12-2021)

14. Levels of insulin-like growth factor in bovine, goat, and sheep milk in different lactation periods: The etiological factor of cancer in humans

Marian Simonov, Vasyl Vlizlo, Volodymyr Stybel, Ruslan Peleno, Volodymyr Salata, Taras Matviishyn, Mariia Кhimych and Oleksii Gorobei

International Journal of One Health, 7(2): 246-250

ABSTRACT

Background and Aim: Despite the positive aspects of consuming dairy products, there are also some reservations. Recently, a large number of publications have demonstrated convincing evidence of a relationship between the intake of dairy products and the development of cancer in humans. This study aimed to determine the level of insulin-like growth factor [IGF], which can cause cancer in consumers, in the milk of farm animals (cows, goats, and sheep) obtained at different stages of lactation.

Materials and Methods: Enzyme-linked immunosorbent assay and statistical method were used for data processing. The IGF level in milk was determined using an enzyme-linked immunosorbent assay. Milk was collected from cows of Holstein and Ukrainian Black-and-White dairy breed, goats of Alpine and Saanen breed, and sheep of Tsigai and East Friesian breed. Milk samples in cows were collected at the beginning, on the peak, and at the end of lactation.

Results: The obtained results showed that the highest IGF level was detected in milk obtained at the beginning of lactation, but its level depended on the animal species. More specifically, the highest level of the hormone was typical for goat's milk and for cow's milk of Holstein breed. Slightly lower values were revealed in the milk of cows of Black-and-White Ukrainian breed and sheep. During lactation, the absolute content of IGF in the milk of all studied animal species decreased. It should be noted that the most pronounced decrease in the IGF level in milk during lactation was noted in sheep (25-40 times), and minor changes were observed in goats (12-14 times). The results also showed that there were individual variations in the IGF level in milk in farm animals.

Conclusion: The highest IGF level was noted in goat's milk, and the lowest level was observed in sheep's milk. Cow's milk, in this regard, had an intermediate IGF level. The level of this particular hormone depends on the period of lactation and the individual characteristics of the animal. It is advisable to include IGF as one of the assessment factors of dairy products' safety because this hormone can cause cancer in consumers of milk and dairy products.

Keywords: cows, goats, insulin-like growth factor, lactation, milk, sheep.

Rift valley fever in Africa with the emerging interest in Libya

Review (Published online: 09-12-2021)

13. Rift valley fever in Africa with the emerging interest in Libya

Abdusalam S. Mahmoud, Osama K. Sawesi, Osama R. El-Waer and Emad M. Bennour

International Journal of One Health, 7(2): 237-245

ABSTRACT

Rift valley fever (RVF) is an acute vector-borne viral zoonotic disease of domestic and wild ruminants. The RVF virus (RVFV) belonging to the Phlebovirus genus of the Bunyaviridae family causes this disease. Studies have shown that mosquitoes are the vectors that transmit RVFV. Specifically, Aedes and Culex mosquito species are among the many vectors of this virus, which affects not only sheep, goats, buffalo, cattle, and camels but also human beings. Since the 30s of the last century, RVF struck Africa, and to a lesser extent, Asian continents, with subsequent episodes of epizootic, epidemic, and sporadic outbreaks. These outbreaks, therefore, resulted in the cumulative loss of thousands of human lives, thereby disrupting the livestock market or only those with seropositive cases. After that outbreak episode, RVF was not reported in Libya until January 13, 2020, where it was reported for the 1st time in a flock of sheep and goats in the southern region of the country. Although insufficient evidence to support RVF clinical cases among the confirmed seropositive animals exists, neither human cases nor death were reported in Libya. Yet, the overtime expansion of RVF kinetics in the Libyan neighborhoods, in addition to the instability and security vacuum experienced in the country, lack of outbreak preparedness, and the availability of suitable climatic and disease vector factors, makes this country a possible future scene candidate for RVF expansion. Urgently, strengthening veterinary services (VS) and laboratory diagnostic capacities, including improvement of monitoring and surveillance activity programs, should be implemented in areas at risk (where imported animals crossing borders from Libyan neighborhoods and competent vectors are found) at national, sub-national, and regional levels. The Libyan government should also implement a tripartite framework (one health approach) among the veterinary public health, public health authority, and environmental sanitation sectors to implement RVF surveillance protocols, along with an active partnership with competent international bodies (OIE, FAO, and WHO). Therefore, this review comprises the most updated data regarding the epidemiological situation of RVF infections and its socioeconomic impacts on African and Asian continents, and also emphasize the emerging interest of RVF in Libya.

Keywords: Africa, epidemiology, Libya, one health, rift valley fever.

Improving the knowledge of high school students about zoonotic diseases from pets in Medellín-Colombia

Research (Published online: 09-12-2021)

3. Improving the knowledge of high school students about zoonotic diseases from pets in Medellín-Colombia

Natalia Uribe Corrales, Karen Velásquez Giraldo, Catalina María Saldarriaga Garcés and Angie Lorena Navarro Giraldo

Veterinary World, 14(12): 3091-3098

ABSTRACT

Background and Aim: The "One Health" concept is a global strategy that recognizes that public health is related to animal health and the environment; however, the role of domestic animals and their involvement in the transmission of zoonoses is often underestimated. The aim of the study was to evaluate and improve the knowledge about zoonotic diseases of domestic animals in high school students from Medellín, Colombia.

Materials and Methods: A quasi-experimental intra-subject study was carried out. This study was conducted with 11th-grade students from four schools in Medellín, Colombia. A structured multiple-choice questionnaire was used from March 2021 to May 2021. The research had two phases, first, "naive" knowledge and learning. Then, descriptive, association, and comparative analysis were carried out using absolute and relative frequencies, Pearson's Chi-square test, and MacNemar's test with a value of p<0.05 considered statistically significant.

Results: A research poll from 315 students of four private schools found that feeding their pets with raw food and leftovers cooked for human consumption were common practices; the results also show a lack of knowledge of their pets' immunization deworming status. It was understood that when the students were able to identify at least two symptoms of zoonoses, one route of its' transmission and two preventive measures, we found that only 12.49% of the polled students had proper knowledge of the disease in domestic animals. After conducting an educational strategy, the findings show a general increase in knowledge, leading us to accept that the academic approach was adequate to statistically increase the recognition of symptoms, routes of transmission and preventive measures (p=0.00).

Conclusion: The use of the theoretical lecture is effective to improve the understanding of the concept of transmission of diseases from animals to humans; in addition, the results show an increase of knowledge in high school students of the related symptoms, transmission routes, and preventive measures of zoonoses diseases in the region.

Keywords: cats, diseases, dogs, education, zoonoses.

Development of a rapid and reliable high-performance liquid chromatography method for determination of water-soluble vitamins in veterinary feed premix

Research (Published online: 08-12-2021)

2. Development of a rapid and reliable high-performance liquid chromatography method for determination of water-soluble vitamins in veterinary feed premix

Md. Zahangir Hosain, S. M. Shariful Islam and Md. Mostofa Kamal

Veterinary World, 14(12): 3084-3090

ABSTRACT

Background and Aim: Determination of trace amounts of vitamins in multi-component feed premix is a troublesome analytical procedure. In this study, a simple and rapid high-performance liquid chromatography (HPLC) method was developed and validated for the concurrent detection and quantitation of four water-soluble vitamins such as thiamine, riboflavin, pyridoxine, and cyanocobalamin in veterinary feed premixes.

Materials and Methods: The chromatographic separation of the vitamins was carried out at 35°C temperature on a reversed-phase C18 column using a gradient pump mode. Mobile phase constituents were solvent (a): 25 mM Potassium dihydrogen phosphate and 5 mM sodium hexanesulfonate in deionized water having pH-4.0 and solvent and (b) 5 mM sodium hexanesulfonate in methanol. Detection was performed with HPLC ultraviolet/visible detection set at 278 and 361 nm wavelength in two different channels. The flow rate was 1.2 mL/min and the total run time was 25 min.

Results: The method was validated according to the International Conference on Harmonization and Food and Drug Administration guidelines and acceptance criteria for system suitability, precision, linearity, and recovery were met in all cases. The relative standard deviation for system suitability and precision was <2% for all vitamins. The linearity of the calibration curves was excellent (R2>0.999) at concentration of 5, 10, 15, 20, 25, and 30 μg/mL for all vitamins. The limits of detection values were 0.0125, 0.0017, 0.0064, and 0.0065 μg/mL for thiamine, riboflavin, pyridoxine, and cyanocobalamin, respectively, and the limits of quantification values were 0.0378, 0.0051, 0.0213, and 0.0198 μg/mL for thiamine, riboflavin, pyridoxine, and cyanocobalamin, respectively. The recovery percentages ranged from 88% to 115%.

Conclusion: The overall parameters of the proposed method met the validation criteria and this method could be a highly desirable technique for routine analysis of water-soluble vitamins in veterinary feed premix.

Keywords: method development, veterinary feed-premix, water-soluble vitamin.