جستجوی مقالات مرتبط با کلیدواژه « culture » در نشریات گروه « دامپزشکی »

Mycoplasma agalactiae (M. agalactiae) is known as the main etiological agent of contagious agalactia (CA). The CA is a disease affecting dairy sheep and goats, the main characteristics of which include keratoconjunctivitis, arthritis, and mastitis. This pathogen results in milk production reduction and suppression, thereby leading to serious economic loss. In the present study, 125 sheep and goat samples were collected from 15 provinces of Iran. Cultural and molecular methods were used for sample characterization. After extracting genomic DNAs using the phenol/chloroform method, the PCR technique was employed to detect Mycoplasma genus in 163bp fragment of 16S rRNA gene (M-PCR) and M. agalactiae in 800bp fragment of conserve and specific P30 lipoprotein gene (P30-PCR) in cultural and clinical samples. Finally, to validate the experimental approach, a 375 bp amplicon of P80 lipoprotein was amplified using the MA-PCR. Out of 125 samples under investigation, 43 cases were positive, and Mycoplasma colonies were observed in the pleuropneumonia-like organisms agar culture. Based on the results of the M-PCR method, 61 specimens (out of 125 samples) were scored positive for Mycoplasma presence. Furthermore, 20 samples were positive according to the P30-PCR data. It should be mentioned that the MA-PCR was performed based on the P80 gene on 125 total samples to furtherverify the results for M.agalactiae detection. Based on the obtained data, P30 and P80 genes were presented and amplified in all Iranian M. agalactiae isolates (n=20). Our results indicated that the P30 gene was conserved and specific to all Iranian M. agalactiae isolates and this new P30-PCR method (as an MA-PCR technique) might be useful in the detection of this pathogen.

Equine salmonellosis is an important infection with a wide variety of consequences including develop-ment of acute salmonellosis in the cases of predisposing factors, nosocomial infections, public health risk, and environmental contaminations.

The aim of this study was to evaluate the fecal shedders of Salmonella spp. in the horses of Kurdistan province of Iran using phenotypic and molecular approach.

A total of 130 fresh feces were randomly collected from horses in four age groups and both sexes in four seasons from all over Kurdistan province. The samples were analyzed for the isolation of Salmonella spp. with culture and biochemical method. An invA-based polymerase chain reaction (PCR) method was also carried out for detection of Salmo-nella spp. in pooled fecal samples, simultaneously. The isolates were further serotyped and the antimicrobial profile of the isolates was determined using Kirby-Bauer method.

The results showed 1.53% (n=2) and 7.69% (n=10) by bacteriological methods and PCR method, respec-tively. There was no significant relation between the frequencies of Salmonella shedders and age, sex and season (p ≥0.05). The two isolates were recognized as Salmonella Typhimurium, showing 100% resistance against ampicillin, tetracycline, streptomycin, sulphamethoxazole, and chloramphenicol, and 50% resistance against gentamycin.

Rapidity and accuracy of PCR versus phenotypic method makes it an appropriate procedure for the surveillance programs regarding Salmonella detection in feces. Approximately high prevalence of subclinical form in equine salmonellosis or Salmonella fecal carriers in the studied region is instigated to seriously apply strategies to manage and control the distribution of infection to susceptible hosts.

Pneumonia due to Mycoplasma infections can cause serious health problems and economic losses in small ruminants industry.

The aim of this study was isolation and identification of Mycoplasmas in sheep naturally infected with pneumonia in Northeastern Iran.

This study used histopathology, culture and polymerase chain reaction (PCR) to examine samples from 50 lungs of sheep naturally infected with Mycoplasmas.

Grossly, irregular consolidation with lobular or lobar to diffuse pattern in the cranioventral to caudal lobes of affected lungs were observed. Histopathologically, bronchointerstitial pneumonia in 38 (76%), and purulent to fibrinopurulent bronchopneumonia in 12 (24%) affected sheep were diagnosed. DNA was extracted from lung tissue samples and replicated using genus and species specific primers for Mycoplasma. Mycoplasma growth was observed in 3 (6%) of a total of 50 lung samples. Genus-specific Mycoplasma DNA was identified by PCR in 12 (24%) of samples. Two (4%) and 7 (14%) samples of these 12 cases were positive for reaction with species-specific primers of Mycoplasma ovipneumoniae and Mycoplasma arginini, respectively.

Our results showed that M. ovipneumoniae and M. arginini were the two agents that can be involved in inducing lung consolidation and pneumonia in sheep and PCR was more successful than the culture in detecting Mycoplasmas.

Mycoplasma gallisepticum and Mycoplasma synoviae are the causative agents of avian mycoplasmosis in commercial poultry. Among the available tools, polymerase chain reaction (PCR) and culture are confirmatory tools for the diagnosis of mycoplasmosis after the initial serological screening of suspected birds. Overall, 181 samples were analyzed, 152 (84%) and 103 (57%) of which were found positive by PCR and culture, respectively. Further, 54 (92%) broiler samples were found positive for general avian mycoplasma. Among the total positive samples, MS positivity was as high as 72 (47%) by PCR, while it was 45 (44%) by culture. MG positivity was 23% and 25% in PCR- and culture-positive samples. MG grows more easily compared to MS. The agreement value between the tests was 67%. Overall, flock wise prevalence was not much varied. The prevalence of mycoplasmosis was higher during winter. Our study confirmed that PCR is the most sensitive and reliable tool for the diagnosis of avian mycoplasmosis in field samples.

Avian influenza H9N2 subtype viruses have had a great impact on Iranian industrial poultry production economy since introduction in the country. To approach Rapid and precise identification of this viruses as control measures in poultry industry, a real time probe base assay was developed to directly detect a specific influenza virus of H9N2 subtype -instead of general detection of Influenza A viruses- which has been endemic over two last decades in the country. An Iranian avian influenza virus strain of A/Iran/chicken/772/1998 H9N2 subtype were selected as reference strain for of primers and probe designing. The high agreement value of 99% indicated that the devolved real time assay for detection of H9 subtype viruses could easily replace the conventional method of virus isolation particularly in investigation of viruses like national surveillance plan. The limit of detection was almost one EID50 which was the least real infectious unit could be detected. So it can be said that this sensitive assay provided a powerful tool to not to miss any significant viral biological activity neither in the host body nor in the environment. A high level of correlation coefficient (R2 = 0.998) also indicated a good correlation between Ct values and viral concentrations. , it can be conclude that the real time RT-PCR could be easily replace virus isolation in detection of H9N2 influenza viruses especially in large monitoring program. The ability in quantifying of the virus concentration extends usage of test in more accurate studies.

L’agalaxie est une maladie infectieuse et contagieuse chez les petits ruminants engendrée notamment par le Mycoplasmaagalactiae (M. agalactiae). Différents facteurs contribuent à l’apparition del’agalaxie , mais M. agalactiae a été reconnu comme lun des agents étiologiques les plus importants. Dans cetteétude, lincidence de M. agalactiaeà l‘origine de lagalaxie contagieuse a été évaluéedans le Sud-est de lIran. Des prélèvements ont été menés en respectant les consignes de lorganisation vétérinaire iranienne sur le lait, la conjonctive, leslésionsauculairesainsi que sur la la ponction du liquide synovial des troupeaux de brebis et chèvres suspectés d’être contaminés. La présence de Mycoplasma et de M. agalactiae a été détectée en utilisant les techniques de culture microbienne et de PCR. Mycoplasma et M. agalactiae ont été identifiés par PCR via l’amplification de la séquence 16S-rRNA desgènes cibles et par la caractérisation des lipoprotéines. Le taux de contamination par le genre Mycoplasmaa été respectivement estimé a14.8% et 36.0% par culture et par PCR. Les infections causées par M. agalactiaeconstituées 6.1% des échantillons. Cette étude souligne l’importance didentifier dautres espèces de Mycoplasma dans les échantillons de petits ruminants engagé avec maladie de l’agalaxie contagieuse.

Mycoplasma capricolumsubsp. capricolumest lun des agents infectieux responsable de lagalactie contagieuse chez les brebis et les chèvres dans les pays de la Méditerranée et du Moyen-Orient. Mycoplasma agalactia, Mycoplasma mycoidessubsp. capri et Mycoplasma capricolum subsp.capricolum sont les agents pathogènes de l’agalactie contagieuse. L’objectif de cetteétude étaitl’isolement et l’identification de Mycoplasma capricolumsubsp.capricolumchez lesbrebis soupçonnées d'être atteintsd’agalactie contagieuse dans la province de lAzerbaidjan oriental en Iran. Un total de 272 échantillons, comprenant74 échantillons de lait, 72 prélèvements oculaires et 126 prélèvements auriculaires ont été prélevés à partir de 49 brebis soupçonnées d'être contaminées par l’Agalactie contagieuse. Tous les échantillons ont été testés simultanément par des méthodes de culture et moléculaire. Dans la méthode moléculaire, tous les cas de mycoplasmes positifs ont été examinés en termes d’appartenance aux espèces M. mycoides et M.capricolum subsp. capricolum. Sur un total de 272 échantillons, 67 échantillons ont été identifiés positifs par la méthode de culture, 87 échantillonsse sont avérés positifs par la méthode de réaction en chaîne par polymérase et les deux méthodes ont montré que 62 échantillons étaient positifs aux mycoplasmes . M. mycoides a été identifié dans 2 prélèvements oculaires, 2 prélèvements auriculaires et 1 échantillon de lait. L’espèce M ;capricolumsubsp. capricolum a été isolée et identifiée à partir de 4 échantillons positifs de M. mycoides. Cet article représentele premier rapport soulignant la présence de Mycoplasma capricolum subsp.capricolum chez les brebis dela province d’Azerbaidjan oriental. Les résultats de cette étude montrentque les prélèvements oculaires, les prélèvements auriculaires et les échantillons de lait sont appropriés pour lisolement et lidentification de M. capricolum subsp.capricolum.

Identification of exogenous factors affecting spermatogonial stem cells (SSCs) proliferation in vitro, provides worthy ways to study the basic biology of the cells. The aim of this study was to investigate the effects of a GnRH analogue (alareline acetate) on SSCs colonization in short-term co-culture with sertoli cells. Five, three-month old Holstein male calves were used to isolate spermatogonial and sertoli cells. Testicular germ cell collection was made by enzymatic digestion methods. The cells were co-cultured in a 15 day period and in vitro effects of various doses (0.5, 1, 2 and 4 µg/ml) of GnRHa on SSCs colonization were assessed. Effects of GnRHa on SSCs proliferation were dose dependent. In conclusion, it was demonstrated that 1 µg/ml GnRHa was the optimum dose for SSCs colonization in comparison with control group. The highest treatment dose (4 µg/ml GnRHa), negatively affected SSCs colonization in comparison with control group.

Mycoplasma capricolum subsp capricolum (Mcc) is one of the etiological agents of contagious agalactia(C.A) in goats which can cause significant economic losses. The aim of this study was to detect Mcc from goats of Qom province in Iran. A total of 111 samples were collected from suspected goats to C.A and cultured in PPLO broth supplemented for Mcc isolation. The bacteria DNAs were extracted from clinical samples and the PCR assay was performed to detect Mycoplasma genus, cluster Mycoides and Mcc from culture. Out of the 111 samples, 33(29.7%) sample cultures were shown positive and typical Mycoplasma colonies in PPLO agar culture method, 53 (47.7%) samples were scored positive by Mycoplasma genus PCR, 8 (7%) of the samples were scored positive by using myciodes cluster PCR and finally 2(1.8%) of the samples were scored positive by using Mcc PCR method. The result of this study showed that Mcc was detected for the first time from Qom province in Iran. Therefore, Qom could be one of the geographical distribution and efficient factors of Mcc in Iranian goats. The lung samples and lymph nodes also could be significant samples for detection of Mcc.

Mycoplasma agalactiae (M. agalactiae) is one of the main causes of contagious agalactia, an infectious syndrome of sheep and goats in Khuzestan province –southwest of Iran that is characterized by mastitis and subsequent failure of milk production, arthritis, abortion and keratoconjunctivitis. This study was carried out to isolation and identification of M. agalactiae with culture and polymerase chain reaction (PCR) method from sheep in Khuzestan province, Iran. A total of 91 samples were collected from milk secretion, eye, ear, nose and joint exudates of sheep. All samples were cultured in PPLO broth supplemented for isolation of M. agalaciae. Extraction of the DNA of bacteria was done by phenol/chloroform method and the PCR assay was applied for detection of Mycoplasma genus in 163bp fragment of 16S rRNA gene and M. agalactiae in 375bp fragment of lipoprotein gene from culture as same as in clinical samples. Out of the 91 samples, 34(37.36%) cultures were shown positive and typical Mycoplasma colonies in PPLO agar culture diagnostic method and 47(51.65%) were scored positive by Mycoplasma genus PCR, 8(8.79%) of the samples were scored positive by using M. agalactiae PCR as diagnostic method. Out of the 91 samples, 26 samples were shown both positive in the culture and PCR, 5 samples were shown both positive in the culture, MPCR and MAPCR. 15 samples were negative in the culture and positive in PCR whereas only 3 samples were positive in culture and negative in PCR. The results showed that the more isolations of M. agalactiae were taken from eye and less in ear and nose samples. M. agalactiae was one of the main factors of contagious agalactia that was detected for the first time from sheep in Khuzestan province.

Contagious agalactia (C. A) is an infectious syndrome of sheep that is characterized by mastitis and subsequent failure of milk production، arthritis، abortion and keratoconjunctivitis. Mycoplasma agalactiae (M. agalactiae) is the main cause of the disease in sheep. The aim of this study was isolation and identification of M. agalactiae with culture and polymerase chain reaction (PCR) assay from sheep of Qom province in Iran. A total of 102 samples were collected from milk secretion، eye، ear and joint exudates of sheep. All samples were cultured in PPLO broth supplemented for M. agalaciae isolation. The bacteria DNAs were extracted by phenol/chloroform method and the PCR assay was applied for detecting of Mycoplasma genus in 163bp fragment of 16S rRNA gene and M. agalactiae in 375bp fragment of lipoprotein gene from culture as same as in clinical samples. Out of the 102 samples، 19 (18. 63%) cultures were shown positive and typical Mycoplasma colonies in PPLO agar culture diagnostic method and 59 (57. 8%) were scored positive by Mycoplasma genus PCR، 19 (18. 62%) of the samples were scored positive by using M. agalactiae PCR as diagnostic method. Out of the 102 samples، 19 samples were shown both positive in the culture and PCR، 42 samples were shown both negative in the culture and PCR. 40 samples were negative in the culture and positive in PCR whereas only one sample was positive in culture and negative in PCR. The results showed that the more isolations of M. agalactiae were taken from milk and less in joint samples. M. agalactiae was one of the main factors of contagious agalactia that was detected for the first time from sheep in Qom province.