Agar Gel Diffusion Test Research Articles

Prevalence of selected viral infections in various age groups of village chickens in Borno State, Nigeria

A preliminary survey for antibodies against some selected viruses in village chickens of various age groups was carried out in Borno State, Nigeria. Three hundred and twenty samples consisting of 160 sera from adult chickens, 60 sera from chicks and 100 yolk samples from embryonated en eggs were tested against Newcastle disease (ND), influenza (FLU) A and B viruses, and egg drop syndrome (EDS) - 76 viruses using haemagglutination inhibition (HI) test; and infectious bursal disease (IBD) virus using the agar gel diffusion test (AGDT). Eighty-five percent of the samples tested positive to one or more viruses, with IBD having the highest percentage prevalence (45.7%) followed by FLU A (26.5%), ND (22.4%), EDS-76 (19.3%) and FLU B (0%). The percentage of mixed infectious among different age groups was highest among chicks. The importance of village chickens in the spread and maintenance of viral diseases of poultry are discussed from an epidemiological point of view.

MICROSCOPICAL AND SEROLOGICAL STUDIES WITH ULTRASTRUCTURE DESCRIPTION OF SARCOCYSTIS SPECIES IN SHEEP IN ASSIUT

This study was conducted to determine theinfection rate of Sarcocystis species in sheep in Assiut. A total of 100 slaughtered sheep examined for Sarcocystis, no macroscopic cysts were observed, 93% (93/100) were positive by microscopical examination and 28% (28/100) were positive by using Agar gel diffusion test.Theinfection rate of Sarcocystis had very high significant differences between microscopic examination and Agar gel diffusion test. High statistical significant effect was found on the infection rate of Sarcocystis in different examined muscles of sheep. The highest infection rate was recorded in oesophagus (71%), followed by diaphragm (65%), tongue (58%), skeletal muscles (53%) and heart muscles (43%). The infection rate of Sarcocystis in males was 95.2% (40/42) while in females were 91.4% (53/58) by microscopical examination. Higher infection rate of Sarcocystis were detected in sheep 6 months - 2 years age 95% (38/40) than that of sheep between 2-4 years old 92.7% (38/41) and those equal or older than 4 years 89.5% (17/19). No significant difference between infection and age groups or sex of animals was observed. The use of transmission electron microscope (TEM) allowed the identification of S. tenella and S. arieticanis in sheep in Assiut. Examination of S. tenella cyst wall with TEM showed palisade-like villar protrusions while, examination of S. arieticanis cyst wall showed hair-like villar protrusions parallel to cyst wall.

Partial biological and molecular characterization of a Cucumber mosaic virus isolate naturally infecting Cucumis melo in Iran.

Melon seedlings showing systemic chlorotic spots and mosaic symptoms were collected in central part of Iran, and a virus was isolated from diseased plants by mechanical inoculation. The virus systemically infected the most inoculated test plants by inducing mosaic symptoms, while, in the members of Fabaceae family and Chenopodium quinoa induced local lesions. Agar gel diffusion test using a polyclonal antiserum against a squash Cucumber mosaic virus (CMV) isolate showed the presence of CMV in the mechanically inoculated plants (designated CMV-Me). The virus was purified by polyethylene glycol precipitation and differential centrifugation. A polyclonal antiserum was produced against the virus that reacted specifically with virus antigen in ELISA and agar gel diffusion tests. The virus was molecularly characterized by PCR amplification of the full length of the coat protein gene using cucumovirus genus specific primer pair CPTALL-3/CPTALL-5 and sequence analysis of the resulting product. No RNA satellite was detected using the primer pair CMVsat3H/sat5T7P. Phylogenetic analysis based on the coat protein amino acid sequences showed that CMV-Me belongs to Subgroup IB. These results may be helpful in melon breeding programs, focusing on plant resistance to plant viruses including CMV.

Serotyping reanalysis of unserotypable Actinobacillus pleuropneumoniae isolates by agar gel diffusion test.

We observed increasing unserotypable (UT) Actinobacillus pleuropneumoniae isolates using agar gel diffusion (AGD) test. To reanalyze their serovar, we performed rapid slide agglutination (RSA) test and multiplex PCR for 47 UT isolates. Of these, 25 were serovar 1 (UT-serovar 1), 20 were serovar 2 (UT-serovar 2) and 2 were serovar 15 (UT-serovar 15). We examined serotyping antigen extraction temperature to determine heat influence. UT-serovar 1 and 15 were influenced by heat, because their precipitation lines were observed in the case of low antigen extraction temperature. To investigate the relationship between antigenicity and genotype, we performed pulsed-field gel electrophoresis (PFGE) analysis using UT-serovar 2 and 15. The predominant PFGE pattern of UT-serovar 2 was identical to that of serovar 2.

PARASITOLOGICAL AND SEROLOGICAL STUDY ON FASCIOLA DIAGNOSIS IN CATTLE AND BUFFALOES IN ASSIUT GOVERNORATE

Received at: 30/3/2014 Accepted: 16/4/2014 The present study was carried out to investigate the prevalence of fascioliasis among 100 cattle and buffaloes of different age and sex in Assiut Governorate. Samples were examined by microscopic examination, Agar Gel Diffusion Test AGDT and ELISA techniques to detect Fasciola species eggs and antibodies, respectively. Prevalence of Fasciola was 8% by microscopic examination, 32% by AGDT and 60.86% by ELISA test. Cattle showed higher prevalence of infection 61.4% than buffaloes 29.4%. Female cattle and buffaloes were more susceptible to the infection 64.55% than male 23.8%. Fasciola was more frequently recorded in 2-4 years old cattle and buffaloes 76% followed by more than 4 years 46.7% then less than 2 years of age 20%. All positive results with fecal egg examination and AGDT were also positive with ELISA. Serological examination by AGDT and ELISA confirms the microscopical examination. The results of this study revealed that the agar gel diffusion test and ELISA could become a useful tools to diagnose fascioliasis in cattle and buffaloes.

Haemophilus parasuisSerovars Isolated from Pathological Samples in Northern Italy

From January 2007 to December 2011, a total of 106 Haemophilus parasuis strains isolated from pigs were serotyped by agar gel diffusion test (DG). Serovar 4 was the most prevalent (24.5%), followed by serovar 13 (19.8%) and serovar 5 (11.3%). Twenty-nine strains were non-typeable (27.3%). The strains were divided into two groups, depending on whether they were isolated from specific pathological lesions of systemic disease such as polyserositis, arthritis or meningitis (73 cases of 106) or from the lower respiratory tract of pigs suffering from bronchopneumonia (33 cases of 106). Serovars 4 and 13 had a higher prevalence in systemic infection (polyserositis) than in respiratory disease only. Pasteurella multocida (14/106), Streptococcus suis (7/106), Actinobacillus pleuropneumoniae (4/106), Bordetella bronchiseptica (3/106) and Arcanobacterium pyogenes (3/106) were isolated in association with H. parasuis.

Replication and attenuation of foot and mouth disease virus in chick embryo

This study includes replication and attenuation of foot and mouth disease virus type O which isolated from infected calves. Many passages for the virus in chick-Embryo were established as a substitute method to the tissue culture which is highly caustic in contrast to the chick embryo. The virus passed ten consequent passages which lead to the reduce of the titer of the virus from 106.53 TCID50/ 0.1 ml in cattle testis tissue culture to 103 TCID50/ 0.1 ml. the pathogenecity of attenuated FMD virus were also studied in both chick-embryo and guinea pigs. Using agar gel diffusion test precipitation antibodies was detected in guinea pig serum after 14 and 21 days post exposure to the attenuated virus. The inoculated guinea pig group with the chick-embryo attenuated virus appear resistance to the challenge virus. The result suggested the efficacy of attenuation of foot and mouth disease virus by using chick-embryo system for immunization against this disease on the level of laboratory Animal.

Replication and attenuation of foot and mouth disease virus in chick embryo

This study includes replication and attenuation of foot and mouth disease virus type O which isolated from infected calves. Many passages for the virus in chick-Embryo were established as a substitute method to the tissue culture which is highly caustic in contrast to the chick embryo. The virus passed ten consequent passages which lead to the reduce of the titer of the virus from 106.53 TCID50/ 0.1 ml in cattle testis tissue culture to 103 TCID50/ 0.1 ml. the pathogenecity of attenuated FMD virus were also studied in both chick-embryo and guinea pigs. Using agar gel diffusion test precipitation antibodies was detected in guinea pig serum after 14 and 21 days post exposure to the attenuated virus. The inoculated guinea pig group with the chick-embryo attenuated virus appear resistance to the challenge virus. The result suggested the efficacy of attenuation of foot and mouth disease virus by using chick-embryo system for immunization against this disease on the level of laboratory Animal.

An outbreak of peste des petits ruminants (PPR) in camels in the Sudan

In mid-August 2004, an outbreak of a previously unknown fatal disease of camels was reported to Kassala State veterinary authorities. Several areas in the state were visited during August–October 2004 to collect epidemiological data and specimens for diagnosis. Clinically the disease was characterized by sudden death of apparently healthy animals and yellowish and later bloody diarrhea and abortion. The disease outbreaks coincided with the seasonal movement of animals towards autumn green pasture. Death was always sudden and proceeded with colic and difficulty in respiration. Mortality rate ranged between 0% and 50% and vary in accordance with the area with a mean of 7.4%. More than 80% of deaths were in pregnant and recently-delivered she-camels. All age, sex and breed groups were affected but more than 50% of deaths were reported in adult animals in comparison to calves and young camels. The main post-mortem findings include lung congestion and consolidation, paleness and fragility of liver, enlarged lymph nodes and congestion and hemorrhage of small intestine and stomach. Agar gel diffusion test (AGDT), RT-PCR and virus isolation in cell culture gave positive results for peste des petits ruminants virus (PPRV), a virus belonging to the Morbillivirus, Genus, member of the family Paramyxoviridae. The effect of this new devastating disease on camel production in the affected area was discussed as well as proposals for future research.

Purification of anatid herpesvirus 1 particles by tangential-flow ultrafiltration and sucrose gradient ultracentrifugation

Anatid herpesvirus 1 (AHV-1) infection causes substantial economic losses to the world-wide waterfowl production. However, little is known about the efficient method used to study the purification of AHV-1 and the negative staining morphology of the purified virus particles. This lack of knowledge is one of the important factors that have affected the progress of research studies on AHV-1 molecular virology to such an extent that they are lagging far behind those on other members of the same family Herpesviridae. Therefore, an efficient method for purifying AHV-1 from cell-culture medium has been developed. Abundant AHV-1 particles, whose morphological features match those of herpesvirus, were obtained by using the following procedures: (1) conventional differential centrifugation for removal of debris after cell disruption, (2) tangential-flow ultrafiltration coupled with sucrose density gradient ultracentrifugation for isolation of the virus, and (3) conventional differential ultracentrifugation for virus concentration. The purified AHV-1 particles were subjected to transmission electron microscopy (TEM), infectivity and recovery tests, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), immunoblotting assay, and agar gel diffusion test (AGDT). The results of examinations revealed that purified AHV-1 particles were free of visible contamination or degradation. The purified AHV-1 particles were biologically active and were successful in initiating infection upon inoculation into susceptible duck embryo fibroblast. The procedures are reliable technically and feasible for purification of large volumes of viruses.

Investigation Of Rota Virus In Dogs

397 fecal samples were collected from dogs of different breeds, ages andsex. Detection of rotavirus was achieved by two techniques latexagglutination test and agar gel diffusion test. Blood samples werecollected to study various hematological parameters.The results showed that 95 samples were positive to rotavirus 23samples (24.2%) were from diarrheic cases and 72 samples (75.8%)were from non diarrheic cases.Younger dogs were more affected with rotavirus infection 17 positivecases than older ages 6 positive cases. The results showed that diarrheiccases caused by rotavirus were 23 cases (25.3%) that were less than thediarrheic cases caused by other agents 68 cases (74.7%).It had been shown that the ages less than one year (11 cases) were moreaffected with rotavirus and complication with dehydration than agesolder than one year (2 cases).It was found that the morbidity rate in males (53%) was significantlyhigher than females (47%).The Terrier breed showed a high morbidity rate 42% than others Mixed,Greatdane, Goledretriver and Doberman.Our study showed that latex agglutination test was more sensitive thanagar gel diffusion test and the positive samples out of 200 samples weredetected by using latex agglutination test were 39 samples while 18positive samples by agar gel diffusion test.There were no differences in blood parameters of the dogs that showedpositive results of rotavirus in comparing them with the negativeresults.The study showed that rotavirus is widely spreaded among doges withgastrointestinal diseases, it showed that latex agglutination test is morereliable for quick clinical diagnosis.

Comparative study of hydropericardium hepatitis syndrome vaccine in poultry

Two commercial types of Hydro pericardium Hepatitis Syndrome (HHS) oily inactivated vaccines were evaluated. The first was (Angavac) French in origin (Merial Co.) & the 2nd is (Nobilis Hepatitis) Holland in origin (Intervet Co.). A unified program of vaccination had been used for both vaccines. One & ten days old chicks were vaccinated by subcutaneous & intramuscular route of injection with both vaccines in each age separately . Serum antibodies titer weekly measured by using Indirect haemagglutination test (IHA) & agar gel diffusion test. Challenge test at 4 week of age was done by virulent avian adeno virus serotype – 4 (VAAV-4) as well as virulent field isolate was used as challenge virus. The result showed highest antibodies titer at week age. Angavac vaccine gave highest titer than Nobilis Hepatitis vaccine & IHA was more sensitive than AGDT. All groups vaccinated with Angavac gave 100% protection in challenge test with (VAAV-4) at 28 days. While in Holland vaccine the protection ratio was 60-80% only, but the ratio of protection of both vaccines did not exceed than 40-50% when the field isolate was used as challenge virus.

Evaluación de desinfectantes de amonio cuaternario sobre cepas bacterianas de origen animal

The authors evaluate two quaternary ammonium compounds, first and second generation, using three methods: minimum bactericide concentration, the suspension test of the German Society for Hygiene and Microbiology and the agar gel diffusion test. The compounds were tested against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Enterococcus faecalis from sheep and pigs. The results show that quaternary ammonia compounds with dodecyl chains have greater efficacy than benzalconium chloride, and that quaternary ammonium compounds are more effective against gram-positive bacteria. No resistance linked to the strains used in the tests was detected. Finally, the authors demonstrate that all three techniques are valid for the evaluation of quaternary ammonium disinfectants.

Application of Agar Gel Diffusion Test to the Diagnosis of Fasciolosis in Cattle and Buffaloes in the Red River Delta of Vietnam

The agar gel diffusion test (AGDT) was applied to detect Fasciola infection in several regions of Vietnam where equipment for fecal examination such as a microscope is not readily available. A 1% solution of agar dissolved in 10% saline, and the antigen prepared from adult individuals of Fasciola sp. were used for the test. The test was specific for Fasciola infection, which was detected in the sera from about 50% of the cattle and water buffaloes examined. The gel and antigen can be stored in a refrigerator for more than 3 months and used for the test. The results of this study revealed that the agar gel diffusion test could become a useful tool to diagnose fasciolosis in cattle and buffaloes at the Regional Animal Health Centers in Vietnam.

Antigen-induced production of interferon-γ in samples of peripheral lymph nodes from sheep experimentally inoculated with Mycobacterium avium subsp. paratuberculosis

The production of interferon-γ (IFN-γ) in response to Johnin purified protein derivate was measured in samples of the prescapular lymph node (PLN) from 10 sheep, aged 2 years, and nine sheep, aged 1 year that had been inoculated orally with Mycobacterium avium subsp. paratuberculosis within their first month of life. Ten non-inoculated sheep, aged 1 year, constituted the negative control group. The results obtained in the PLN IFN-γ assay were compared with those derived from serological tests: a complement fixation test (CFT), agar gel diffusion test (AGID) and enzyme-linked immunosorbent assay (ELISA), as well as an IFN-γ test on samples of blood. Among the 19 inoculated sheep, 16 gave positive reactions in the PLN IFN-γ assay on samples incubated overnight, and 18 tested positive when the assay was applied to PLN samples incubated for 48 h. In comparison, three, four and seven inoculated sheep gave positive reactions in the ELISA, CFT and in the blood IFN-γ assay on samples incubated overnight, respectively. The AGID and IFN-γ assay on blood samples incubated for 48 h detected eight inoculated animals. Twelve inoculated sheep, that tested positive in the PLN IFN-γ assay were clinically normal, gave negative results in an IS900-based polymerase chain reaction (PCR) assay on samples of ileum and ileocaecal lymph node and had no histological evidence of paratuberculosis, but tested positive on more than two occasions in sequential serological testing before necropsy. None of the 10 non-inoculated sheep tested positive in the AGID, CFT, ELISA, blood IFN-γ assay on samples incubated overnight and for 48 h or the PLN IFN-γ assay on samples incubated overnight, but one gave a positive result in the PLN IFN-γ assay on samples stimulated for 48 h. It is likely that the positive reactions obtained by the PLN IFN-γ assay in the 12 inoculated sheep that tested negative in the PCR assay and histopathological examination represents immunological evidence of latent infection or previous exposure to M. paratuberculosis rather than active infection.

In vitro antibiotic sensitivity of strains of Ornithobacterium rhinotracheale isolated in the Netherlands between 1996 and 1999

All the strains of Omithobacterium rhinotracheale isolated in the Netherlands between 1996 and 1999 were tested in the agar gel diffusion test for their sensitivity to amoxycillin, tetracycline, enrofloxacin and...

Molecular characterization of a tombusvirus associated with a disease of statice (Goniolimon tataricum (L.) Boiss.).

A tombusvirus was isolated from plants of statice (Goniolimon tataricum) showing malformation, mosaic and necrosis of the leaves and poor flowering. Electron microscopy of infected tissues revealed the presence of vesiculated structures derived from altered peroxisomes. In Northern blots four major RNA species were detected, corresponding to viral genomic, subgenomic and defective interfering (DI) RNAs. Both genomic and DI RNAs were cloned and sequenced, and shown to have structure and organization similar to those of viruses of the genus Tombusvirus. In agar gel diffusion tests, spurs were formed when the statice isolate was compared with the cherry strain but not with the BS3 strain of Tomato bushy stunt virus.

HYDROPERICARDIUM IN BROILER CHICKENS IN IRAQ: ISOLATION AND IDENTIFICATION OF THE CAUSATIVE VIRUS

An adenovirus was successfully isolated from infected liver, spleen and pericardial fluid from infected broiler chicks in primary cell cultures prepared from chicken embryo liver and kidneys. The isolated virus induced cytopathogenic effect (CPE) in infected cell cultures which consisted of cell granulation, aggregation and rounding of infectedd cells. An adenovirus like morphology was identified by Electron microscope examination. In addition the isolated virus was diagnosed, by agar gel diffusion test (AGD). Indirect immune fluorescenct test (IFA) and serum neutralization (SN). Using reference hyperimmune serum and sera from recovered chicks

The Radish Mosaic Virus—A New Virus for Bulgaria

ABSTRACTIn one plant Raphanus raphanistrum with mosaic symptoms the Radish mosaic virus (RaMV) was established for the first time in Bulgaria. This virus was identified by biological test, indexes for the stability of the sap “in vitro” serological agar-gel diffusion test and tests for transmission by aphids.

Characteristics of a Cowpea Chlorotic Mottle Virus Isolate from Nigeria

Cowpea chlorotic mottle virus (CCMV, bromovirus group) was isolated from Desmodium heterocarpon and Clitoria ternatea plants growing in experimental plots at the International Institute of Tropical Agriculture, Ibadan, Nigeria. It was identified by serology, particle morphology, pH sensitivity, and analyses of viral coat protein and nucleic acids. In reciprocal agar-gel diffusion tests, the CCMV-Nigerian isolate and the type CCMV strain were found to be serologically distinct. This is the first report of the occurrence of a virus resembling CCMV outside of the United States and Costa Rica