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Genetic relationship of Salmonella isolates found in subcutaneous abscesses in leopard geckos (Eublepharis macularius)
2020
Zając, Magdalena | Maluta, Aleksandra | Wasyl, Dariusz | Skarżyńska, Magdalena | Lalak, Anna | Samcik, Ilona | Kwit, Renata | Szulowski, Krzysztof
The article describes the occurrence and phylogenetic relationship of Salmonella isolates found in subcutaneous abscesses of leopard geckos. The aim of the study was to determine the cause of the abscesses and to characterise isolated Salmonella strains. Samples of abscesses from five animals and internal organs (lungs, liver, and gut) of three of them were tested for Salmonella according to the PN-EN ISO 6579:2002/A1:2007 standard. The antimicrobial resistance was evaluated by minimal inhibitory concentrations and the genetic similarity of the isolates was assessed with pulsed field gel electrophoresis (PFGE). In total, seventeen Salmonella isolates belonging to five different serovars were found to be susceptible to all tested antimicrobials except streptomycin. The serovars were S. Hadar, S. Fluntern, S. Tennessee, S. enterica subsp. salamae 55:k:z₃₉, and S. Kentucky. Up to three serovars from different organs were isolated from the same individual. In two geckos, Salmonella were detected in the lungs. In three serovars, XbaI-PFGE typing revealed indistinguishable isolates from organs and abscesses. Multiple Salmonella serovars might be involved in abscess formation and infections. The occurrence of the same PFGE profiles of the isolates may testify to the role of opportunistic organisms in causing infection.
Afficher plus [+] Moins [-]Occurrence of Marek’s disease in Poland on the basis of diagnostic examination in 2015–2018
2020
Marek’s disease (MD) is a tumourous disease caused by Marek’s disease virus (MDV) and most commonly described in poultry. The aim of the study was to determine the occurrence of Marek’s disease virus infections in Poland and analyse clinical cases in the years 2015–2018. The birds for diagnostic examination originated from 71 poultry flocks of various types of production. Birds were subjected to anatomopathological examination post mortem, during which liver and spleen sections and other pathologically changed internal organs were taken. These sections were homogenised with generally accepted methods, then total DNA was isolated and amplified with a real-time PCR. A pair of primers complementary to the MDV genome region encoding the meq gene were used. MDV infection was found predominantly in broiler chicken flocks (69.01%), and also in layer breeder (9.85%) and commercial layer flocks (7.04% each). The results of research conducted in the years 2015–2018 clearly indicate that the problem of MDV infections is still current.
Afficher plus [+] Moins [-]Point-of-service diagnostic technology for detection of swine viral diseases
2020
Nannucci, Lapo | Barattini, Paolo | Bossis, Ioannis | Woźniakowski, Grzegorz | Balka, Gyula | Pugliese, Carolina
A research project is underway aiming to develop a field diagnostic tool for six important viruses of the pig sector, namely: African swine fever virus (ASFV), porcine reproductive and respiratory syndrome virus (PRRSV), swine influenza virus (SIV), porcine parvovirus (PPV), porcine circovirus (PCV2), and classical swine fever virus (CSFV). To obtain a preliminary sounding of the interest in this type of instrument among its potential operators, a questionnaire was drawn up and submitted to three categories of stakeholders: farmers, veterinarians, and others (including scientific and technical staff working on animal farms). Four countries participated: Italy, Greece, Hungary, and Poland. In total, 83 replies were collected and analysed in a breakdown by stakeholder type and pertinence, where the areas were the importance of the main diseases within the different countries, diagnostic tool operational issues, and economic issues. The main end-users of this kind of instrument are expected to be private veterinarians and pig producers. The infectious agents seeming to be most interesting to diagnose with the instrument are PRRSV, SIV, PPV, and PCV2. The most decisive parameters which have been selected by the stakeholders are sensitivity, cost, simplicity, and time required to obtain results. The economic issue analysis showed that the majority of those who would prefer to buy rather than rent the device are willing to pay up to €3,000 for a diagnostic field tool.
Afficher plus [+] Moins [-]A novel, rapid, and simple PMA-qPCR method for detection and counting of viable Brucella organisms
2020
Zhang, Shi-Jun | Wang, Lu-Lu | Lu, Shi-Ying | Hu, Pan | Li, Yan-Song | Zhang, Ying | Chang, Heng-Zhen | Zhai, Fei-Fei | Liu, Zeng-Shan | Li, Zhao-Hui | Ren, Hong-Lin
The plate counting method widely used at present to discern viable from non-viable Brucella in the host or cell is time-consuming and laborious. Therefore, it is necessary to establish a rapid, simple method for detecting and counting viable Brucella organisms. Using propidium monoazide (PMA) to inhibit amplification of DNA from dead Brucella, a novel, rapid PMA-quantitative PCR (PMA-qPCR) detection method for counting viable Brucella was established. The standard recombinant plasmid with the target BCSP31 gene fragment inserted was constructed for drawing a standard curve. The reaction conditions were optimised, and the sensitivity, specificity, and repeatability were analysed. The optimal exposure time and working concentration of PMA were 10 min and 15 μg/mL, respectively. The correlation coefficient (R²) of the standard curve was 0.999. The sensitivity of the method was 10³ CFU/mL, moreover, its specificity and repeatability also met the requirements. The concentration of B. suis measured by the PMA-qPCR did not differ significantly from that measured by the plate counting method, and the concentrations of viable bacteria in infected cells determined by the two methods were of the same order of magnitude. In this study, a rapid and simple PMA-qPCR counting method for viable Brucella was established, which will facilitate related research.
Afficher plus [+] Moins [-]Potential determinants of Clostridium spp. occurrence in Polish silage
2020
Goldsztejn, Magdalena | Grenda, Tomasz | Kozieł, Nina | Sapała, Magdalena | Mazur, Małgorzata | Sieradzki, Zbigniew | Król, Beata | Kwiatek, Krzysztof
Silage quality deteriorates with Clostridium spp. contamination, and if consumed, such silage jeopardises herd health and productivity. Minimising its occurrence reduces economic and animal welfare risks. The study investigated the influence of environmental and technological determinants on the Clostridium genus’ occurrence in silage. Analyses were conducted on 305 silage samples directly collected from farms located in all Polish provinces. Cultures and isolates were evaluated phenotypically and examined for occurrence of Clostridium spp., particularly C. perfringens and C. botulinum using PCR techniques. The results were statistically analysed using the ᵡ² test for continuous and Student’s t-test for non-continuous values. The most influential effect on Clostridium spp. occurrence is exerted by factors potentially associated with primary production, like the type of fertilisation and the contamination level of the ensiled feed material. Clostridium spp. was detected in 232 (76%) samples, and C. perfringens strains, predominantly toxinotype A, in 79 (26%). C. botulinum occurrence was not detected. Deterioration of silage by clostridia could be prevented by a properly conducted ensiling process with the addition of starter cultures, but the presence of spores mainly depends on primary production and the extent of contamination of the feed material.
Afficher plus [+] Moins [-]Effect of bathing in a 0.1% aqueous solution of ethacridine lactate on selected physiological parameters of Cornu aspersum Müller edible snails
2020
Ziętek, Jerzy | Dzięgiel, Beata | Wójcik, Alicja | Wilczyńska, Anna | Adaszek, Łukasz | Winiarczyk, Stanisław
The prohibition of antibiotic use in edible snails obligates breeders to treat bacterial infections by different means, of which a common one is a bath in Gram-positive– and partially Gram-negative–bactericidal ethacridine lactate solution. The aim of the study was to determine the effect of bathing Cornu aspersum Müller snails in a 0.1% aqueous solution of ethacridine lactate on selected physiological parameters of haemolymph. The study included 80 snails, divided into two equal groups (study and control). The study group was subjected to bathing in ethacridine lactate and the control group to bathing in tap water. Both groups were treated daily for seven days. The number of haemocytes in the haemolymph, the activity of alanine (ALT) and aspartate (AST) aminotransferases, and the concentration of urea were determined. In the study group, after exposure to ethacridine lactate solution an increase in ALT activity, changes in the De Ritis ratio, an increase in the amount of haemocytes, and a decrease in body weight were found. No such changes were detected in the control group snails or in animals after the first bath. Multiple applications of a 0.1% ethacridine lactate bath may adversely affect Cornu aspersum Müller snails.
Afficher plus [+] Moins [-]Metagenomic analysis of acquired antibiotic resistance determinants in the gut microbiota of wild boars (Sus scrofa) – preliminary results
2020
Libisch, Balázs | Keresztény, Tibor | Kerényi, Zoltán | Kocsis, Róbert | Sipos, Rita | Papp, Péter P. | Olasz, Ferenc
Land application of manure that contains antibiotics and resistant bacteria may facilitate the establishment of an environmental reservoir of antibiotic-resistant microbes, promoting their dissemination into agricultural and natural habitats. The main objective of this study was to search for acquired antibiotic resistance determinants in the gut microbiota of wild boar populations living in natural habitats. Gastrointestinal samples of free-living wild boars were collected in the Zemplén Mountains in Hungary and were characterised by culture-based, metagenomic, and molecular microbiological methods. Bioinformatic analysis of the faecal microbiome of a hunted wild boar from Japan was used for comparative studies. Also, shotgun metagenomic sequencing data of two untreated sewage wastewater samples from North Pest (Hungary) from 2016 were analysed by bioinformatic methods. Minimum spanning tree diagrams for seven-gene MLST profiles of 104 E. coli strains isolated in Europe from wild boars and domestic pigs were generated in Enterobase. In the ileum of a diarrhoeic boar, a dominant E. coli O112ab:H2 strain with intermediate resistance to gentamicin, tobramycin, and amikacin was identified, displaying sequence type ST388 and harbouring the EAST1 toxin astA gene. Metagenomic analyses of the colon and rectum digesta revealed the presence of the tetQ, tetW, tetO, and mefA antibiotic resistance genes that were also detected in the gut microbiome of four other wild boars from the mountains. Furthermore, the tetQ and cfxA genes were identified in the faecal microbiome of a hunted wild boar from Japan. The gastrointestinal microbiota of the free-living wild boars examined in this study carried acquired antibiotic resistance determinants that are highly prevalent among domestic livestock populations.
Afficher plus [+] Moins [-]Redescription of Quasiamidostomum fulicae (Rudolphi, 1819) Lomakin, 1991 (Nematoda: Amidostomatidae), a parasite of Fulica atra (Gruiformes)
2020
Królaczyk, Katarzyna | Zaborski, Daniel | Dzierzba, Emil | Kavetska, Katarzyna M.
Quasiamidostomum fulicae (Rudolphi, 1819) Lomakin, 1991, is a species of which the systematic position is still unclear, and it is reported in the literature under many synonyms. In the present study, an attempt has been made at establishing the ultimate systematic position of Quasiamidostomum fulicae against the backdrop of selected Amidostomatinae species. The parasites were identified based on measurements of external and internal structures. Ecological analysis of Q. fulicae was carried out using the quantitative indices (frequency, prevalence, mean intensity, relative abundance, and dominance index). Statistical analyses (discriminant analysis) were performed on measurement data. The intestines of 77 coots were examined. They yielded a total of 398 parasites, including 67 identified as Q. fulicae. Both males and females were located in the muscular gizzard. The morphometric analysis of Q. fulicae in this study showed the dimensions of all the internal organs to be in agreement with measurements reported by other authors. The discriminant analysis, used to find the differences between the examined nematode species (Amidostomoides acutum, A. petrovi, A. monodon, Amidostomum anseris, and Quasiamidostomum fulicae), gave highly significant results (P < 0.0001) with respect to both males and females. The results justify the separation of Q. fulicae from the genus Amidostomum.
Afficher plus [+] Moins [-]Snail eggs as a raw material for the production of a caviar substitute
2020
Maćkowiak-Dryka, Monika | Szkucik, Krzysztof | Pyz-Łukasik, Renata
Snail eggs can be the raw material for the production of a caviar substitute. The substitute varies from the original in caloric value and nutrient content which determine the nutritional value of every foodstuff. The present study aimed to determine and compare the nutritional value and protein quality of eggs from two subspecies of edible snail. The chemical composition of the snail eggs i.e. Cornu aspersum maxima and Cornu aspersum aspersum was determined in accordance with international standards. In order to evaluate the protein quality of the eggs of the two studied snail subspecies, the chemical score (CS), and a reference protein were used. Significant differences in the content of water, ash, and carbohydrates, but comparable protein and fat contents and caloric values were found. The protein in the eggs of the snails was complete by the measure of the model adopted for this study, however, meeting the daily essential amino acid requirements of an adult would require an immense supply of both species’ eggs. Snail eggs of the Cornu genus were characterised by much lower nutritional value in comparison with caviar and caviar substitutes.
Afficher plus [+] Moins [-]Genetic characterisation of African swine fever virus in outbreaks in Ha Nam province, Red River Delta Region of Vietnam, and activity of antimicrobial products against virus infection in contaminated feed
2020
Tran, Ha Thi Thanh | Truong, Anh Duc | Ly, Duc Viet | Vũ, Thị Hảo | Hoàng, Văn Tuấn | Nguyễn, Thị Chính | Chu, Thi Nhu | Nguyen, Thi Huyen | Pham, Ngoc Thi | Nguyen, Tinh | Yersin, Andrew G. | Dang, Hoang Vu
African swine fever (ASF) was officially reported in Vietnam in February 2019 and spread across the whole country, affecting all 63 provinces and cities. In this study, ASF virus (ASFV) VN/Pig/HaNam/2019 (VN/Pig/HN/19) strain was isolated in primary porcine alveolar macrophage (PAM) cells from a sample originating from an outbreak farm in Vietnam’s Red River Delta region. The isolate was characterised using the haemadsorption (HAD) test, real-time PCR, and sequencing. The activity of antimicrobial feed products was evaluated via a contaminated ASFV feed assay. Phylogenetic analysis of the viral p72 and EP402R genes placed VN/Pig/HN/19 in genotype II and serogroup 8 and related it closely to Eastern European and Chinese strains. Infectious titres of the virus propagated in primary PAMs were 10⁶ HAD₅₀/ml. Our study reports the activity against ASFV VN/Pig/HN/19 strain of antimicrobial Sal CURB RM E Liquid, F2 Dry and K2 Liquid. Our feed assay findings suggest that the antimicrobial RM E Liquid has a strong effect against ASFV replication. These results suggest that among the Sal CURB products, the antimicrobial RM E Liquid may have the most potential as a mitigant feed additive for ASFV infection. Therefore, further studies on the use of antimicrobial Sal CURB RM E Liquid in vivo are required. Our study demonstrates the threat of ASFV and emphasises the need to control and eradicate it in Vietnam by multiple measures.
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