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In-house validation method for quantification of amoxicillin in medicated feedingstuffs with the use of HPLC-DAD technique Полный текст
2020
Patyra, Ewelina | Kwiatek, Krzysztof
A high-performance liquid chromatographic–diode array detector (HPLC-DAD) method for the determination of amoxicillin in medicated feedingstuffs was developed and validated. The method was used to investigate the quality requirements of animal feedingstuffs (declared content of active substance and feed homogeneity). Two-gram samples were extracted by potassium phosphate buffer solution. Extracts were filtered and directly analysed by HPLC-DAD without further clean-up. Amoxicillin was separated by acetonitrile and 0.01M phosphate buffer (pH 5.0) on a Phenomenex Luna C18 column. This method provided average recoveries of 76.1 to 81.6% with coefficients of variation (CV, %) for repeatability and reproducibility in the ranges of 3.7–7.2% and 5.3–7.6%, respectively. The limit of detection was 51.2 mg/kg and limit of quantification was 103.0 mg/kg. The method was successfully validated and proved to be efficient, precise, and useful for quantification of amoxicillin in medicated feedingstuffs.
Показать больше [+] Меньше [-]Seroprevalence of antibodies against Anaplasma phagocytophilum and Borrelia burgdorferi in horses (Equus caballus) from northern Algeria Полный текст
2020
Laamari, Abdelouahab | Azzag, Naouelle | Tennah, Safia | Derdour, Salima-Yamina | China, Bernard | Bouabdallah, Ryhan | Ghalmi, Farida
Horses (Equus caballus) are susceptible to tick-borne diseases. Two of them, Lyme borreliosis due to Borrelia burgdorferi and granulocytic anaplasmosis due to Anaplasma phagocytophilum were investigated in Algerian horses. The diseases have been less extensively studied in horses and results pertinent to Algeria have not been published. Blood samples were obtained from 128 horses. IgG antibodies directed against Anaplasma phagocytophilum and Borrelia burgdorferi were detected by an indirect immunofluorescence antibody test (IFAT) and ELISA. The potential effects of age, gender, breed, and health status on seropositivity were also evaluated. Using IFAT, 28 (21.8%) and 25 (19.5%) animals were positive for B. burgdorferi and A. phagocytophilum, respectively. Using ELISA, 19 (14.8%) and 33 (25.9%) animals were positive for these bacteria. The study shows that horses in Algeria are exposed or co-exposed to tick-transmitted zoonotic bacterial species.
Показать больше [+] Меньше [-]Meta-analysis of genetic diversity of the VP1 gene among the circulating O, A, and SAT2 serotypes and vaccine strains of FMD virus in Egypt Полный текст
2020
El Nahas, Abeer F. | Salem, Sayed A.H.
Three strains of the FMD virus (A, O, and SAT 2) were recognised as causes of the FMD circulating in Egypt. The aims of this study were to trace the FMDV isolates from outbreaks in Egypt to understand their epidemiology and evolution and to understand the situation of the vaccine strains compared with the circulating serotypes. A meta-analysis was carried out by using the data available for FMD outbreaks in Egypt from GenBank and the World Reference Laboratory for Foot-and-Mouth Disease (WRLFMD); a comparison was done with both data sets for the three serotypes. MEGA-X was used for the evolution analysis, through constructions of phylogenetic trees for all sequences recorded in GenBank for each serotype in different Egyptian outbreaks in different years and also within the same year. Additionally, nucleotide substitution rate, molecular clock, and mean evolutionary rates were estimated for the three serotypes to understand and compare their evolution. Absence of some records of certain serotype outbreaks from the WRLFMD database was noted as were subsequent missing appropriate vaccine programmes. Genetic variation was recorded among the virus isolates within the same years and also the vaccine strain was associated with up to 26 amino acid substitutions. The evolution rate of the SAT2 strain was the highest of the circulating strains. SAT2 had high amino acid substitution per year at an important immunogenic site (130–170), serotype A had less, and serotype O the least. The need for different strategies for vaccine serotype selection is indicated.
Показать больше [+] Меньше [-]Occurrence of Marek’s disease in Poland on the basis of diagnostic examination in 2015–2018 Полный текст
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.
Показать больше [+] Меньше [-]Effect of goose parvovirus and duck circovirus coinfection in ducks Полный текст
2020
Liu, Jie | Yang, Xiaoxia | Hao, Xiaojing | Feng, Yongsheng | Zhang, Yuli | Cheng, Ziqiang
Coinfection of goose parvovirus (GPV) and duck circovirus (DuCV) occurs commonly in field cases of short beak and dwarfism syndrome (SBDS). However, whether there is synergism between the two viruses in replication and pathogenicity remains undetermined. We established a coinfection model of GPV and DuCV in Cherry Valley ducks. Tissue samples were examined histopathologically. The viral loads in tissues were detected by qPCR, and the distribution of the virus in tissues was detected by immunohistochemistry (IHC). Coinfection of GPV and DuCV significantly inhibited growth and development of ducks, and caused atrophy and pallor of the immune organs and necrosis of the liver. GPV and DuCV synergistically amplified pathogenicity in coinfected ducks. In the early stage of infection, viral loads of both pathogens in coinfected ducks were significantly lower than those in monoinfected ducks (P < 0.05). With the development of the infection process, GPV and DuCV loads in coinfected ducks were significantly higher than those in monoinfected ducks (P < 0.05). Extended viral distribution in the liver, kidney, duodenum, spleen, and bursa of Fabricius was consistent with the viral load increases in GPV and DuCV coinfected ducks. These results indicate that GPV and DuCV synergistically potentiate their replication and pathogenicity in coinfected ducks.
Показать больше [+] Меньше [-]Animal coronaviruses in the light of COVID-19 Полный текст
2020
Domańska-Blicharz, Katarzyna | Woźniakowski, Grzegorz | Konopka, Bogdan | Niemczuk, Krzysztof | Welz, Mirosław | Rola, Jerzy | Socha, Wojciech | Orłowska, Anna | Antas, Marta | Śmietanka, Krzysztof | Cuvelier-Mizak, Beata
Coronaviruses are extremely susceptible to genetic changes due to the characteristic features of the genome structure, life cycle and environmental pressure. Their remarkable variability means that they can infect many different species of animals and cause different disease symptoms. Moreover, in some situations, coronaviruses might be transmitted across species. Although they are commonly found in farm, companion and wild animals, causing clinical and sometimes serious signs resulting in significant economic losses, not all of them have been classified by the World Organization for Animal Health (OIE) as hazardous and included on the list of notifiable diseases. Currently, only three diseases caused by coronaviruses are on the OIE list of notifiable terrestrial and aquatic animal diseases. However, none of these three entails any administrative measures. The emergence of the SARS-CoV-2 infections that have caused the COVID-19 pandemic in humans has proved that the occurrence and variability of coronaviruses is highly underestimated in the animal reservoir and reminded us of the critical importance of the One Health approach. Therefore, domestic and wild animals should be intensively monitored, both to broaden our knowledge of the viruses circulating among them and to understand the mechanisms of the emergence of viruses of relevance to animal and human health.
Показать больше [+] Меньше [-]Colistin resistance of non-pathogenic strains of Escherichia coli occurring as natural intestinal flora in broiler chickens treated and not treated with colistin sulphate Полный текст
2020
Majewski, Michał | Łukomska, Anna | Wilczyński, Jarosław | Wystalska, Danuta | Racewicz, Przemysław | Nowacka-Woszuk, Joanna | Pszczola, Marcin | Anusz, Krzysztof
A significant threat to public health is presented by antibiotic-resistant strains of bacteria, selective pressure on which results from antibiotic use. Colistin is an antibiotic commonly used in veterinary medicine, but also one of last resort in human medicine. Since the 2015 discovery in China of the mcr-1 gene encoding colistin resistance in Enterobacteriaceae, other countries have noted its presence. This study was to find the mcr-1 gene prevalence in E. coli isolated from poultry slaughtered in Poland. Cloacal swabs were taken from December 2017 to October 2018 from broiler chickens in three regions. The samples (n = 158) were grouped as flocks treated with colistin sulphate (n = 87) and those not treated (n = 71). Resistance to antimicrobials commonly used in poultry was evaluated by minimum inhibitory concentration. The presence of the mcr-1 gene was confirmed by PCR. Isolates containing the mcr-1 gene were yielded by 11.27% of the samples from not treated flocks and 19.54% of those from treated flocks, but no statistically significant difference in the prevalence of the gene was seen between the groups. The results clearly preclude intensification of selective pressure for colistin resistance due to colistin sulphate treatment because they show that the avian gastrointestinal tract was already inhabited by colistin-resistant E. coli by the time the chickens came to the poultry house.
Показать больше [+] Меньше [-]Cloning and identification of PK15 cells for enhanced replication of classical swine fever virus Полный текст
2020
Yin, Mei | Hu, Dongfang | Li, Peng | Kong, Lingyun | Ning, Hongmei | Yue, Feng | Jiang, Jinqing | Wang, Xuannian
Classical swine fever virus (CSFV) causes an economically important and highly contagious disease of pigs, leading to economic losses around the world. Attenuated live vaccines with CSFV antigens have played an important role in the prevention and control of the disease. Porcine kidney 15 (PK15) cells have been widely used for the propagation of CSFV, but this cell line is not efficient or homogeneously susceptible to viral infection. To achieve a homogeneous PK15 cell line which enabled high titre replication of CSFV, we used the limiting dilution cell cloning method. We developed two cell clones, PK15-1A6 and PK15-3B1, which respectively have high- and low-permissive phenotypes to CSFV infection. The PK15-1A6, PK15-3B1, and PK15 parent cells showed different characteristics in cell proliferation rate, susceptibility to CSFV infection, and CSFV production. The mean virus titres per millilitre reflected by TCID₅₀ values in PK15-1A6, PK15-3B1, and PK15 parent cells were 106.85, 103.63, and 104.74, respectively. The PK15-1A6 cell clone is more permissive to CSFV infection than the PK15 parent cells. The screened high-permissive cells will be useful for CSFV propagation and vaccine development in vitro, and facilitate research on the pathogenicity of CSFV.
Показать больше [+] Меньше [-]Evaluation of the effect of surgical and immunological castration of male pigs on boar taint compounds in oral fluid and fat tissue by LC-MS/MS method Полный текст
2020
Woźniak-Sobczak, Barbara | Cybulski, Piotr | Jabłoński, Artur | Witek, Sebastian | Matraszek-Żuchowska, Iwona
An effective way of preventing undesirable boar taint in pork meat caused by the presence of androstenone, skatole and indole is surgical castration of piglets. This, however, arouses growing social opposition. An alternative method of inhibiting the development of unpleasant odour is immune castration. The aim of the study was to compare the effectiveness of both methods of castration for the elimination of the compounds responsible and to assess the suitability of oral fluid for pre-slaughter predictive testing for boar taint. The research material was pooled oral fluid and fat samples taken from gilts and surgically and immunologically castrated piglets. The samples were tested with a liquid chromatography– tandem mass spectrometry method developed in this research. The compounds giving rise to boar taint were found only sporadically above the accepted limits; only one sample of oral fluid contained skatole at a concentration above 200 μg L⁻¹ and one contained indole more concentrated than 100 μg L⁻¹. Indole above the limit value was also detected in one fat sample. In none of the tested samples was androstenone found. The results indicate the similar effectiveness of both methods of piglet castration on the reduction of compounds generating boar taint. The usefulness of testing oral fluid for the ante-mortem prediction of boar taint has not been fully confirmed and further investigation is needed.
Показать больше [+] Меньше [-]Therapeutic effects of revascularisation on the healing of free bone grafts in dogs Полный текст
2020
Zheng Jia-San | Ruan Hong-Ri | Shuang-Qiu | Jing-Nie | Hou Kai-Wen | Rui-Wu
Therapeutic effects of revascularisation on the healing of free bone grafts in dogs Полный текст
2020
Zheng Jia-San | Ruan Hong-Ri | Shuang-Qiu | Jing-Nie | Hou Kai-Wen | Rui-Wu
The therapeutic effect of subcutaneous embedding and revascularisation on the repair of canine bone defects caused by open fracture was examined.
Показать больше [+] Меньше [-]Therapeutic effects of revascularisation on the healing of free bone grafts in dogs Полный текст
2020
Zheng, Jia-San | Ruan, Hong-Ri | Shuang-Qiu, | Jing-Nie, | Hou, Kai-Wen | Rui-Wu,
The therapeutic effect of subcutaneous embedding and revascularisation on the repair of canine bone defects caused by open fracture was examined. A total of 12 adult beagle dogs were randomly split into a control group (group C) and a test group (group T). A section of the radius was removed from each dog under general anaesthesia and the deficit supported by an orthopaedic implant. Group T had the section surgically implanted next to the blood vessel–rich saphenous vein and Group C had it cryopreserved at −80°C. After eight weeks, the bone was surgically implanted back into the matching radial deficit. Bone healing was evaluated by gross morphological and X-ray examinations, post-mortem histology, and successive blood measurements of key bone biochemical markers. At 12 weeks, the bone healing boundary was disappearing more quickly in group T dogs than in their group C counterparts. X-ray and histological examinations showed that the cortical repair of group T subjects was complete and the bony plate arrangement was more regular than that in group C. The levels of bone biochemical markers also proved that the healing state of group T was better. The results showed that the degree of healing, osteoclast activity, and bone formation status of group T were better than those of group C, proving that the vascularised bone graft had a significantly shorter healing time than the cryopreserved bone graft.
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