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Molecular surveillance of gamma coronaviruses in pigeon flocks, Tehran province, 2014-2015
2017
Ghalyanchi Langeroudi, Arash | Karimi, Vahid | Abdi Haji, Mohammad Reza | Vasfi Marandi, Mahdi | Hashemzadeh, Masoud | Maghssoudloo, Hosein | Madhi, Ali
BACKGROUND: Coronaviruses have a wide range of host tropism causing respiratory, enteric and central nervous system diseases in pigs, cats, dogs, rodents, cattle, avian species and human. Coronaviruses undergo genetic mutations and recombination at high rates which make them able to infect a wide range of host species from different geographical locations. According to the possible existence of gammacoronavirus in Iranian bird population including pigeons and lack of information about virus prevalence, isolation and molecular characterization of pigeon coronaviruses are needed. OBJECTIVES: The present study was conducted to detect avian coronavirus in flocks of Tehran province. METHODS: samples were randomly collected from 25 pigeon flocks of Tehran province (250 tracheal swabs and 250 cloacal swabs, 20 specimens from each flock) between 2014-2015. The viral RNA was extracted from swab samples and RT-PCR reaction was run using the QIAGEN one-step RT-PCR Kit with primers targeting nucleocapsid (N) gene and 3’ untranslated region (3’-UTR) of gammacoronavirus. RESULTS: Gammacoronavirus was detected in one out of 25 (4%) flocks. CONCLUSIONS: The results of this study approve the presence of gammacoronaviruses in pigeon population and help to complete the map of epidemiology of the virus in Iran. According to the low prevalence rate of coronavirus in pigeons, samples should be collected from pigeons showing respiratory or enteric signs of disease or from pigeons having contact with other birds or those which are housed near poultry farms.
اظهر المزيد [+] اقل [-]Detection of testosterone residues in farm fish tissue
2009
H. R. Abdel-Dayem | Hanaa M. Soltan | G. Naser
A total of one hundred samples of marketed fish farm; 25each of tilapia (Oreochromis niloticus), Mugil cephalus, Carp (Silver) and fry; were collected from different localities at Al- Behera and Kafr El-Shakh Provinces. The samples were examined for detection of testosterone hormone by Thin Layer chromatography (TLC) as well as it was quantitatively assayed by Radio-Immuno-Assay (RIA). The obtained result revealed that the testosterone residues were detected in flesh of farm fish of Tilapia (Oreochromis nilotocus) and Carp (Silver), each constituting 24% and 4 %; respectively but it could be failed to detect in each of Mugil cephalus and fry. The hormonal residues of testosterone was ranged from 3.25 to 34.9 ng/g with mean value 4.22±1.1 ng/g in positive samples of Tilapia, while only one sample of Carp showed 22.0 ng/g, Mugil cephalus and Fry showed no detectable level of hormonal residues. The public health significance of detectable levels was discussed as well as the recommendations to avoid health hazards from such fish were mentioned.
اظهر المزيد [+] اقل [-]Detection of Chlamydophila abortus in sheep by Polymerase Chain Reaction
2007
Wafaa A. Osman
This study was carried out on 180 pregnant ewes located at Ras Sedr research station - desert research center, from 2003 to 2005. Twenty five cases of abortion were recorded and examined serologically by complement fixation test (CFT). 17 (68%) out of these 25, showed positive results against Chlamydophila abortus and 3(15%) out of 20 apparently healthy pregnant ewes were serologically positive. Due to the fact that both clinical signs and pathological findings are not specific in case of chlamydial infection and also due to the fact that CFT is accompanied by false positive reactions due to cross reactivity between chlamydial species, five samples from serologically positive aborted dams were subjected to polymerase chain reaction (PCR). They revealed positive results for Chlamydophila abortus at 119 bp. Therefore, PCR is proven to be reliable, rapid and specific diagnostic technique in thediagnosis of chlamydial infection.
اظهر المزيد [+] اقل [-]Detection of residues of some heavy metals in fish fillets in Egyptian markets
2010
R. H. Abdel-Dayem | H. M. Soltan | N. M. Marzouk
Fifteen samples of fish fillets were collected from different Egyptian markets to detect the residues of some heavy metals in them. The average weights of fish fillets samples were 250g. The samples were separately packed in polyethylene bag and sent to the laboratory for preparation and analysis. All equipment surfaces and utensils which used in this study were thoroughly cleaned with detergent, rinsed with water, dried and then rinsed with 2-propanol solution. The samples were stored at –2C prior to analysis. The mean levels of lead and cadmium in fish fillets collected from the Egyptian markets were 1.51 0.34 and 0.60 0.0.25 mg/kg; respectively. The concentration of lead in fish muscle were less than the permissible heavy metal limit in fish meat (2.0 mg kg) as stated by the World Health Organization, also these results were in line with the corresponding EU regulations. However the level of cadmium concentration was over than the heavy metal limits in fish meat (0.5 mg/kg) for the World Health Organization.
اظهر المزيد [+] اقل [-]Detection of Chlamydophila psittaci in chickens by complement fixation test and polymerase chain reaction
2007
Wafaa A. Osman | A. L. El-Naggar | Azza S. A. Gooda | Mona A. Mahmoud
This study was carried out on 68 randomly collected chickens located at Ras Sedr Research Station, Desert Research Center, 68 serum samples were examined serologically by complement fixation test (CFT). Twenty out of 68 (29.91%) had antibodies against Chlamydophila psittaci . Ten blood samples of the serologically positive cases were subjected to polymerase chain reaction (PCR) and showed positive results for Chlamydophila psittaci at 119 bp. Therefore PCR was found to be reliable, rapid, sensitive and specific technique for the detection Chlamydophila psittaci in birds. Serologically positive birds did not show any clinical symptoms of disease, but they were in contact with sheep and goat that showed previous abortion and were positive for C. abortus. It is recommended to avoid breeding of chickens with other animal species in the same yard because chickens become asymptomatic carrier with shedding of Chlamydophila psittaci in their feaces and respiratory discharges.
اظهر المزيد [+] اقل [-]Preparation of diagnostic ELISA kits for detection of camelpox virus
2005
E. A. Aboul Soud | Magda, M. Sayed | A. A. Badawi | M. M. Taha | Elham A. El-Ebiary | A. M. Daoud
Peroxidase labeled immunoglobulins to camelpox virus (CPV) were prepared for use in various techniques of ELISA. Ten rabbits and three goats were inoculated with a mixture of camelpox virusand Freund’s adjuvant. Sera were pooled separately on the 10th day post the last inoculation and immunoglobulins were precipitated using saturated ammonium sulphate. The globulins were 2.8 g/dl and 2.5 g/dl for rabbits and goats respectively and used for peroxidase conjugation. The peroxidase labeled immunoglobulins were titrated and evaluated using direct solid phase ELISA, double antibody sandwich ELISA and dot immunoblot ELISA. The prepared conjugates gave specific and clear positive reactions till the dilution of 2000 and 1500 for rabbits and goats immunoglobulins respectively. The prepared labeled immunoglobulins could be successfully used in detection of camel pox viral antigen of local virulent and standard vaccinal strain of the virus using various ELISA techniques.
اظهر المزيد [+] اقل [-]Oestrus synchronisation with progesterone-containing sponge and equine chorionic gonadotropin in Pirlak ewes during the non-breeding season: can Toryum improve fertility parameters?
2020
Kuru, Mushap | Boga Kuru, Buket | Sogukpinar, Osman | Cebi Sen, Cigdem | Oral, Hasan | Kirmizibayrak, Turgut
The aim of the study was to determine the effect of the vitamins, omega-3 polyunsaturated fatty acid and minerals in the supplement Toryum administered before and during oestrus synchronisation on some fertility parameters of ewes during the non-breeding season. The experimental animals were clinically healthy Pirlak ewes, 55–75 days postpartum, aged 2–4 years and weighing 40–50 kg. A sponge was inserted into the vagina for 10 d (G1, n = 30; G2, n = 30) or 14 d (G3, n = 30; G4, n = 30) for oestrus synchronisation, and on the day of removal, 400 IU equine chorionic gonadotropin was injected. Toryum soft capsules were administered individually (1 capsule/ewe p.o.) to G1 and G3 ewes seven days before the sponge was inserted and on the day it was removed. Oestrus detection was started 12 h after sponge removal. Pregnancy was diagnosed by transrectal ultrasonography on the 30ᵗʰ day after mating. The pregnancy rate was statistically different between G1 and G4 (P < 0.05). The onset of oestrus was statistically different (P < 0.001) between the 10-d groups (G1 and G2) and the 14-d groups (G3 and G4). The litter size and oestrus, conception, lambing, multiple birth, and survival rates were not significantly different between the groups (P > 0.05). Toryum administered to Pirlak ewes during progesterone-based oestrus synchronisation protocols during the non-breeding season may increase pregnancy rates. The relationship between Toryum and fertility parameters in ewes would be better understood by comprehensive studies.
اظهر المزيد [+] اقل [-]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.
اظهر المزيد [+] اقل [-]Detection of white sturgeon iridovirus (WSIV) in wild sturgeons (Actinopterygii: Acipenseriformes: Acipenseridae) in Poland
2020
Hofsoe-Oppermann, Paulina | Kiełpińska, Jolanta | Panicz, Remigiusz | Bergmann, Sven M.
White sturgeon iridovirus (WSIV) disease is caused by a virus of the eponymous family and is mostly triggered by stressful environmental conditions, i.e. high rearing density, excessive handling, or temporary loss of water. The aim of this study was to develop the most effective diagnostic method for quick and efficient confirmation or exclusion of the presence of WSIV. A total of 42 samples (spleen, gills, intestine, skin, kidney, and brain) were collected from eight sturgeon (Acipenser gueldenstaedtii and A. oxyrinchus) aged ≤5+ farmed or caught between 2010 and 2014 in open waters (Dąbie Lake and Szczecin Lagoon). They were tested for WSIV presence using conventional PCR, qPCR, and in situ hybridisation (ISH). In gross examination, all fish appeared to be healthy. Neither species showed clinical signs typical of WSIV infection. In the majority of cases, fragments of iridoviral DNA were found using molecular methods in the kidneys, and also in the liver, gills, and skin. The detection rate using ISH was 47.37% and most commonly the brain and kidney tissues were positive. The most efficient of the methods used was real-time PCR, with 100% effectiveness in detection of WSIV DNA. The study demonstrates the capabilities for WSIV diagnosis available to sturgeon farmers and water administrators, indicating useful methods of adequate sensitivity as well as organs to sample in order to achieve the highest probability of viral detection.
اظهر المزيد [+] اقل [-]Occurrence of gastrointestinal parasites in camels in the Tianshan Mountains pastoral area in China
2020
Guowu, Zhang | Kai, Zhang | Xifeng, Wang | Chunhui, Ji | Chengcheng, Ning | Yue, Zhao | Jun, Qiao | Qingling, Meng | Xingxing, Zhang | Kuojun, Cai | Jinsheng, Zhang | Zaichao, Zhang | Xuepeng, Cai
Gastrointestinal parasites are some of the most common pathogens which are seriously harmful to the camel’s health. The infection status of gastrointestinal parasites in camels (Camelus bactrianus) in the Tianshan Mountains pastoral area in China is still unclear. The aim of this study was to investigate the species and infection intensity of gastrointestinal tract parasites in local camels. A total of 362 fresh faecal samples were collected and examined for parasite eggs using the saturated saline floating and natural sedimentation method. The parasite eggs were subjected to morphological and molecular examination and identification, and the infection rate and mean intensity of the parasites were analysed. A total of 15 gastrointestinal tract parasite species’ eggs were identified, with a detection rate of 100%. Ostertagia spp. (100%) and Trichostrongylus spp. (98.1%) were dominant. Camels were often coinfected by 5–14 species. The average number of eggs per gram of faeces was higher for Ostertagia spp. (298), Haemonchus contortus (176) and Nematodirus spp. (138). The number of species of parasites infecting young camels was significantly lower than that of adult camels, but the infection intensity in young camels was significantly higher. Gastrointestinal parasites were highly prevalent in camels from the Tianshan Mountains pastoral area in China. This finding provides important epidemiological data for the prevention and control of associated infections in camels.
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