Two hundred and thirty six semen samples were collected from 120 bulls, 60 buffalo-bulls, and 56 ... more Two hundred and thirty six semen samples were collected from 120 bulls, 60 buffalo-bulls, and 56 rams located on farms of known history of infection with Chlamydophila species. All semen samples were examined by polymerase chain reaction (PCR) and cell culture techniques for detection of Chlamydophila species. The primers were selected to allow the amplification of all target species in a single reaction by identifying conserved sequences in the omp2 gene. PCR assay detected more positive samples (36) from the semen samples collected from different animal species than were detected by the culture method (21). The results indicated that all culture-positive semen samples (21) from different species were PCR positive. The detection limit of the PCR assay was determined with DNA extracted from fourfold serial dilution of C. abortus (B577) and C. pecorum (11/88) cultures and found to be 0.25 inclusion-forming units (IFU) per PCR, while the culture method could not detect less than 4 IFU. This is the first report using PCR for the detection of Chlamydophila species in buffalo-bullsÕ semen and the assay provides a simple, sensitive, rapid, and reliable means for the detection and identification of the organism.
Chlamydophila abortus-DNA was detected using a touchdown enzyme time-release (TETR)-polymerase ch... more Chlamydophila abortus-DNA was detected using a touchdown enzyme time-release (TETR)-polymerase chain reaction (PCR) assay as an improved test for sensitive and rapid diagnosis of abortion in small ruminants. Two hundred and fifty two placentae, liver or spleen tissue samples from aborting ewes and goats or aborted lambs and kids in which C. abortus infection was suspected were examined by TETR-PCR and the results were compared with cell culture. Sixty-five tissue samples were found to be TETR-PCR positive while only 56 samples were cell culture-positive. After resolution of discrepant samples with a confirmatory nested PCR assay, TETR-PCR had a sensitivity of 97% and a specificity of 99.5% while culture had a sensitivity of 84.8% and a specificity of 100%. The analytical sensitivity of the TETR-PCR assay was determined with DNA extracted from 4-fold serial dilution of C. abortus B577 culture and found to be 0.25 inclusion-forming unit per PCR. No reduction in the analytical sensitivity was noted when the assay was tested with mouse liver samples spiked with 4-fold serial dilution of C. abortus B577 culture. No target product was amplified when DNA from Chlamydophila pecorum was tested. TETR-PCR used in this study is a practical, rapid, sensitive and specific assay that could be used for the detection of C. abortus in infected tissue samples. We recommend the use of this assay as a supplemental diagnostic tool for detection of C. abortus in infected tissue samples.
I n this study, culture and RT-PCR assays for detection of fusion protein (F) and glycoprotein ge... more I n this study, culture and RT-PCR assays for detection of fusion protein (F) and glycoprotein genes (G) of bovine respiratory syncytial virus (BRSV) were used in 18 and 15 nasopharyngeal swabs from calves and camels, respectively. Also, the assays were used to test 16 and 13 necropsy samples developing severe respiratory lesions of lung tissues from calves and camels, respectively. The results indicated that, RT-PCR assays more sensitive than the culture method for detection of BRSV in clinical samples. A total of 2, 3 and 5 samples collected from calves were positive by culture, RT-PCR (F) and (G), respectively. On the other hand, 3, 1 and 2 samples collected from camels were positive by culture, RT-PCR (F) and (G), respectively. The results also revealed that the RT-PCR, using G gene as the target, was able to detect BRSV in clinical samples more sensitive than RT-PCR (F). In conclusion, an important application of RT-PCR (G) in the future could be useful for direct, rapid specific and sensitive testing of respiratory secretions, and/or lung tissues from cattle for the presence of BRSV to gather molecular epidemiological information.
3 Abstract: Bovine milk samples were collected from cases of clinical and sub-clinical mastitis, ... more 3 Abstract: Bovine milk samples were collected from cases of clinical and sub-clinical mastitis, respectively and examined bacteriologically and by simplex and multiplex PCR assays for detection of Staphylococcus aureus, Escherichia coli and Streptococcus agalactiae. Escherichia coli was the most common bacteria detected in the collected samples from clinical mastitis cases followed by Staphylococcus aureus and Streptococcus agalactiae; while Staphylococcus aureus was the most common bacteria followed by Escherichia coli and Streptococcus agalactiae in samples collected from sub-clinical mastitis cases. Compared with the cultural method, the simplex and multiplex PCR assays are less time consuming. It took less than 24 hours to be completed, while identification of bacteria to the species levels by conventional microbiological and biochemical methods required more than 72 hours. In conclusion, simplex and multiplex PCR assays can be used as a rapid, sensitive and specific routinely ...
This study was carried out to evaluate, PCR-based method, for detection of cow's milk in wat... more This study was carried out to evaluate, PCR-based method, for detection of cow's milk in wate r buffalo's milk. It utilized primers targeting the mitochondrial 12S rRNA gene. The detection limit of the evaluated PCR method was 0.5% and it was determined using model samples made from buffalo's milk containing defined percentages of cow's milk. The method was also evaluated for its applicability for inspection of 21 market milk samples labeled "buffalo milk". Ten out of the 21 examined milk samples were proven to be pure buffalo's milk; three samples were confirmed to be pure cow's milk while the remaining eight samples were mixed cow and buffalo milk. In conclusion, the PCR assays evaluated in this study can be useful for milk inspection to detect cow's milk in water buffalo milk with a detection limit of 0.5%. Also, analysis of market milk samples revealed that adulteration of buffalo milk by mixing with cow's milk or even substituti...
The occurrence of bovine herpes virus-1 (BHV-1) in camels was studied. A total of 186 pneumonic c... more The occurrence of bovine herpes virus-1 (BHV-1) in camels was studied. A total of 186 pneumonic camel lungs were collected from slaughter houses at four different areas in Sudan during 2000-2006. Using sandwich ELISA 1.6% of 186 tested lungs were found positive for BHV-1 antigen, all were from Tambool at Central Sudan. Direct fluorescent antibody test (FAT) was used to confirm the BHV-1 ELISA positives, all ELISA positives were also positive. PCR was used to detect BHV-1 genome with three positive results. BHV-1 was isolated from two camel lungs in MDBK cells. Isolates were identified using ELISA and FAT. Indirect ELISA was used to detect antibodies to BHV-1 in 260 camel sera; 76.9% were found positive. Highest prevalence was observed in sera from Kordofan (84%) then Blue Nile (80%) and Tambool (76.3%). This is the first report for the detection of BHV-1 antigen, genome using PCR, isolation in cell culture and antibodies in camels in Sudan.
This study was undertaken to investigate the role of parainfluenza virus 3 (PIV3) in respiratory ... more This study was undertaken to investigate the role of parainfluenza virus 3 (PIV3) in respiratory infection of camels. A total of 273 lung specimens from camels with pneumonia lesions were collected from slaughterhouses in four different areas of Sudan. In addition, eight specimens were collected from outbreaks of respiratory infection in camels. Using antigen detection sandwich ELISA kits, six out of the 281 specimens tested were positive for the PIV3 antigen (2.1%); the highest prevalence was noted in Eastern Sudan (4.2%), then in Central and Northern Sudan (1.4%). The direct immunofluorescent test (FAT) was used to confirm the positive reactions for PIV3 by ELISA. The polymerase chain reaction (RT-PCR) was applied for the detection of the PIV3 genome in lungs of camels; two out of four samples which were positive by the PIV3 ELISA were also positive by RT-PCR. Virus isolation was attempted for PIV3 in MDBK cells; four specimens yielded cytopathic virus when inoculated onto the cell culture. The cytopathic effect (CPE) consisted of cell rounding, multinucleated cells, sloughing and elongation of cells, and some syncytia were observed on the 3rd to 7th day post-inoculation. Using commercially available indirect ELISA kits for antibodies to PIV3, 495 camel sera were tested, and the seroprevalence detected was 82.2%. The highest seroprevalence was observed in Central (92.6%), then in Eastern (92.2%) and Central to South Sudan (82.5%); the lowest prevalence was found in Northern Sudan (64.8%).
An evaluation of the polymerase chain reaction (PCR) for detection of Brucella melitensis DNA in ... more An evaluation of the polymerase chain reaction (PCR) for detection of Brucella melitensis DNA in bovine and ovine semen was performed. Since semen contains different components that inhibit PCR ampli®cation, a protocol was used to purify Brucella-DNA from bovine and ovine semen samples prior to conducting ampli®cation of the targeted DNA. When separated fractions of naturally Brucella contaminated semen were analyzed by the PCR, most of B. melitensis DNA were present in the seminal¯uid and non-sperm fractions. The PCR examination results for detection of B. melitensis DNA in different semen fractions were compared with the results for traditional cultural methods of Brucella from semen. The PCR was more sensitive than the traditional cultural methods since it detected Brucella-DNA in 12 (10%) out of 120 semen samples while direct culture detected only 7 (5.8%) in the same semen samples. The limit of detection by PCR was 100 CFU/ml of semen. In addition, the results of PCR were available in one day, whereas isolation and identi®cation of Brucella organisms required days or even weeks. The PCR may be used as a supplementary test for detection of B. melitensis in semen.
This study aimed to investigate the occurrence of respiratory syncytial virus (RSV) infections in... more This study aimed to investigate the occurrence of respiratory syncytial virus (RSV) infections in camels in Sudan. A total of 272 camel lung specimens showing pneumonia were collected from slaughter houses at four different areas in Sudan, additionally 8 specimens were collected from outbreaks of respiratory infection in camels. Using sandwich ELISA kits for RSV antigen detection 4 out of 280 tested lungs (1.4%) were positive, all were from Central Sudan (Tambool slaughter house). FAT was used to confirm the ELISA positives. Polymerase chain reaction RT/PCR was applied for the detection of RSV genome in camel lungs; 1 out of 4 ELISA positives was positive by RT/PCR. Using indirect ELISA kits 135 out of 495 (27.3%) camel sera showed antibodies to RSV, highest prevalence was observed in Western (33.5%) then Central (31.6%) and Eastern Sudan (23.5%). Based on the manufacturer specified calculations for OD readings, most of positive sera (90/135) were low reactive (1+). This is the first report for the detection of RSV antigen, genome and antibody in camels in Sudan.
Trypanosoma evansi is enzootic in camels in Egypt, and water buffaloes act as a reservoir for cam... more Trypanosoma evansi is enzootic in camels in Egypt, and water buffaloes act as a reservoir for camel infection. Molecular techniques have contributed towards understanding the epidemiology of T. evansi. Trypanosoma evansi was detected in acute and chronic stages of the disease in male and female mice by polymerase chain reaction (PCR) using two primers. Two experiments were conducted. In experiment I, two groups consisting of 26 female and 26 male mice received 104 trypanosome by I/P inoculation for each mouse. In experiment II, 42 female and 42 male mice were inoculated I/P with 102 trypanosome/mouse. In addition, five mice were kept as uninfected control for each group. Mice were monitored daily for parasitaemia level during the pre-patent period using the micro-haematocrit centrifugation technique (MHCT) and conventional PCR. The primer pairs, (Trypanosoma brucei) TBR1/2 and TeRoTat1.2 (T. evansi Rode Trypanozoon antigen type [RoTat] 1.2), detected the infection after 24 hours ear...
INTRODUCTION High economic lose was resulting due to the reproductive disorders in cattle and buf... more INTRODUCTION High economic lose was resulting due to the reproductive disorders in cattle and buffaloes such as abortions, repeat breeding and infertility (Yoo et al., 2010). Bovine abortion may be due to many causes, but infectious causes are the most important one (Silva et al., 2009). Worldwide, Brucellosis, Leptospirosis, Mycoplasmosis and Listeriosis are considered serious zoonosis and represent an important cause of reproductive losses in animals, especially in Mediterranean countries and Egypt (Gwida et al., 2016). These diseases are characterized by fast spreading, difficulty of control and prevention, time consuming and the cost of treatment (Selim et al., 2014). As a replacement of time consuming traditional methods, PCR has been used as an important diagnostic tool of abortion in cattle (Anderson, 2007). Several single PCR assays have been used for detection of abortion microbial agents, including Brucella species, pathogenic Leptospira species, Mycoplasma species and Listeria monocytogenes (
THE EGYPTIAN JOURNAL OF EXPERIMENTAL BIOLOGY (Zoology)
Graphene oxide Nano-sheets (GOs) have a wide range of industrial, biochemical and medical applica... more Graphene oxide Nano-sheets (GOs) have a wide range of industrial, biochemical and medical applications regarding their unique physical, chemical and biocompatibility properties. For assessment of toxicological potential of graphene oxide nanosheets ninety male mice were divided into six groups; the control and five treated groups: animals of the control group received an intraperitoneal (i.p) injection of 0.2 ml saline solution (0.9% NaCl) once weekly for eight weeks. The treated groups were received an i.p injection of 10, 50, 100, 250, and 500 µg GOs/kg body weight once weekly, for eight weeks. Animals of all groups were sacrificed after 7, 28, and 56-days post treatment. The present study was conducted to evaluate the genotoxic effects of GOs on mice liver cells using alkali comet assay. In addition, physiological investigations and description of hepatic histopathological alternations were carried out. Results revealed that GOs induced DNA damage (DNA fragmentation), represented in dose and time-dependent increase in % tail DNA migration, tail moment and comet tail length. As well as, a diminish in % head DNA in nuclei of liver of GOs-treated mice versus control groups. The effect of GOs on hepatic superoxide dismutase (SOD), catalase (CAT) activities and glutathione (GSH) level indicated slight decrease at low doses (10 and 50 µg) at all time intervals of experimental period as compared to the control groups. The administration of high doses of GOs (100, 250, and 500 µg) indicated a significant diminish in SOD, CAT activities and decrease in GSH level; with subsequent elevation in malondialdehyde (MDA) levels in liver as compared to control groups. Various hepatic histopathological alternations were dose and time dependent in GO-treated mice versus control. In conclusion, the induction of DNA fragmentation in liver denoting the genotoxicity of GOs in dose and time dependent manner. GOs had the ability for inducing various hepatic alternations in a dose and time dependent manner indicating cytotoxicity presumably by oxidative stress. Thus, GOs should be used under strict control and these serious side effects should be taken into consideration when used in vivo treatments.
Experimental and toxicologic pathology : official journal of the Gesellschaft fur Toxikologische Pathologie, Jan 5, 2017
Graphene and graphene-related materials have broadly applied in biomedical purposes due to their ... more Graphene and graphene-related materials have broadly applied in biomedical purposes due to their unique properties, thus safety evaluation of them is crucial. This study was performed to explore the genotoxic and pulmonary toxic potential of different doses of graphene oxide nanosheets' (GOs) in mice.A total of 90 male mature mice were randomly divided into six groups of fifteen mice per each, five groups were intraperitoneally injected by GO at doses of 10, 50, 100, 250 and 500μg/kg b.w once weekly in addition to the control group that was injected intraperitoneally with 0.2ml saline solution. Five animals from each group were euthanized after 7, 28 and 56days post treatment. Evaluation of genotoxicity was performed through detection of chromosomal aberrations in bone marrow while assessment of lung injury was made by determination of DNA fragmentation in lung specimens using the alkali Comet assay, pulmonary oxidative markers estimation and finally histopathological investigat...
The European journal of contraception & reproductive health care : the official journal of the European Society of Contraception, Jan 20, 2014
Objective To study the association between Porphyromonas gingivalis (P. gingivalis) infection and... more Objective To study the association between Porphyromonas gingivalis (P. gingivalis) infection and recurrent miscarriage. Methods This case control study included women with early pregnancy failure admitted for surgical evacuation of retained products of conception. Cases (group 1) included 50 women with unexplained recurrent early miscarriage whereas the control group (group 2) consisted of 50 women with no such history. The evacuated products of conception, subgingival plaques, cervicovaginal secretions and saliva of all participants were examined to detect P. gingivalis deoxyribonucleic acid (DNA) using a polymerase chain reaction. Results The prevalence of P. gingivalis DNA in the chorionic villous tissue samples of group 1 was significantly higher than in group 2 (8 [16%] vs. 1 [2%], respectively; p = 0.036, odds ratio [OR]: 9.3, 95% confidence interval [CI]: 1.1-76.9). The prevalence of P. gingivalis DNA was significantly higher in cervicovaginal secretions of group 1 than in g...
Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis) is the causative agent of p... more Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis) is the causative agent of paratuberculosis, or Johne's disease, in cattle, with potential involvement in cases of Crohn's disease in humans. Johne's disease is found worldwide and is economically important for both beef and dairy industries. In an effort to characterize this important infection in Egypt, we analysed the ecological and genomic features of recent isolates of M. paratuberculosis. In this report, we examined 26 Holstein dairy herds distributed throughout Egypt, from 2010 to 2013. Using PCR analysis of faecal samples, we estimated a mean herd-level prevalence of 65.4 %, with animallevel infection that reached a mean of 13.6 % among animals suffering from diarrhoea. Whole genome sequencing of field isolates identified numerous single nucleotide polymorphisms among field isolates relative to the standard M. paratuberculosis K10 genome. Interestingly, the virulence of M. paratuberculosis isolates from Egypt revealed diverse virulence phenotypes in the murine model of paratuberculosis, with significant differences in tissue colonization, particularly during the chronic stage of infection. Overall, our analysis confirmed that Johne's disease is a newly identified problem in Egypt and indicated that M. paratuberculosis has potentially diverse genotypes that impact its virulence. Further ecological mapping and genomic analysis of M. paratuberculosis will enhance our understanding of the transmission and evolutionary dynamics of this pathogen under natural field conditions.
Two hundred and thirty six semen samples were collected from 120 bulls, 60 buffalo-bulls, and 56 ... more Two hundred and thirty six semen samples were collected from 120 bulls, 60 buffalo-bulls, and 56 rams located on farms of known history of infection with Chlamydophila species. All semen samples were examined by polymerase chain reaction (PCR) and cell culture techniques for detection of Chlamydophila species. The primers were selected to allow the amplification of all target species in a single reaction by identifying conserved sequences in the omp2 gene. PCR assay detected more positive samples (36) from the semen samples collected from different animal species than were detected by the culture method (21). The results indicated that all culture-positive semen samples (21) from different species were PCR positive. The detection limit of the PCR assay was determined with DNA extracted from fourfold serial dilution of C. abortus (B577) and C. pecorum (11/88) cultures and found to be 0.25 inclusion-forming units (IFU) per PCR, while the culture method could not detect less than 4 IFU. This is the first report using PCR for the detection of Chlamydophila species in buffalo-bullsÕ semen and the assay provides a simple, sensitive, rapid, and reliable means for the detection and identification of the organism.
Chlamydophila abortus-DNA was detected using a touchdown enzyme time-release (TETR)-polymerase ch... more Chlamydophila abortus-DNA was detected using a touchdown enzyme time-release (TETR)-polymerase chain reaction (PCR) assay as an improved test for sensitive and rapid diagnosis of abortion in small ruminants. Two hundred and fifty two placentae, liver or spleen tissue samples from aborting ewes and goats or aborted lambs and kids in which C. abortus infection was suspected were examined by TETR-PCR and the results were compared with cell culture. Sixty-five tissue samples were found to be TETR-PCR positive while only 56 samples were cell culture-positive. After resolution of discrepant samples with a confirmatory nested PCR assay, TETR-PCR had a sensitivity of 97% and a specificity of 99.5% while culture had a sensitivity of 84.8% and a specificity of 100%. The analytical sensitivity of the TETR-PCR assay was determined with DNA extracted from 4-fold serial dilution of C. abortus B577 culture and found to be 0.25 inclusion-forming unit per PCR. No reduction in the analytical sensitivity was noted when the assay was tested with mouse liver samples spiked with 4-fold serial dilution of C. abortus B577 culture. No target product was amplified when DNA from Chlamydophila pecorum was tested. TETR-PCR used in this study is a practical, rapid, sensitive and specific assay that could be used for the detection of C. abortus in infected tissue samples. We recommend the use of this assay as a supplemental diagnostic tool for detection of C. abortus in infected tissue samples.
I n this study, culture and RT-PCR assays for detection of fusion protein (F) and glycoprotein ge... more I n this study, culture and RT-PCR assays for detection of fusion protein (F) and glycoprotein genes (G) of bovine respiratory syncytial virus (BRSV) were used in 18 and 15 nasopharyngeal swabs from calves and camels, respectively. Also, the assays were used to test 16 and 13 necropsy samples developing severe respiratory lesions of lung tissues from calves and camels, respectively. The results indicated that, RT-PCR assays more sensitive than the culture method for detection of BRSV in clinical samples. A total of 2, 3 and 5 samples collected from calves were positive by culture, RT-PCR (F) and (G), respectively. On the other hand, 3, 1 and 2 samples collected from camels were positive by culture, RT-PCR (F) and (G), respectively. The results also revealed that the RT-PCR, using G gene as the target, was able to detect BRSV in clinical samples more sensitive than RT-PCR (F). In conclusion, an important application of RT-PCR (G) in the future could be useful for direct, rapid specific and sensitive testing of respiratory secretions, and/or lung tissues from cattle for the presence of BRSV to gather molecular epidemiological information.
3 Abstract: Bovine milk samples were collected from cases of clinical and sub-clinical mastitis, ... more 3 Abstract: Bovine milk samples were collected from cases of clinical and sub-clinical mastitis, respectively and examined bacteriologically and by simplex and multiplex PCR assays for detection of Staphylococcus aureus, Escherichia coli and Streptococcus agalactiae. Escherichia coli was the most common bacteria detected in the collected samples from clinical mastitis cases followed by Staphylococcus aureus and Streptococcus agalactiae; while Staphylococcus aureus was the most common bacteria followed by Escherichia coli and Streptococcus agalactiae in samples collected from sub-clinical mastitis cases. Compared with the cultural method, the simplex and multiplex PCR assays are less time consuming. It took less than 24 hours to be completed, while identification of bacteria to the species levels by conventional microbiological and biochemical methods required more than 72 hours. In conclusion, simplex and multiplex PCR assays can be used as a rapid, sensitive and specific routinely ...
This study was carried out to evaluate, PCR-based method, for detection of cow's milk in wat... more This study was carried out to evaluate, PCR-based method, for detection of cow's milk in wate r buffalo's milk. It utilized primers targeting the mitochondrial 12S rRNA gene. The detection limit of the evaluated PCR method was 0.5% and it was determined using model samples made from buffalo's milk containing defined percentages of cow's milk. The method was also evaluated for its applicability for inspection of 21 market milk samples labeled "buffalo milk". Ten out of the 21 examined milk samples were proven to be pure buffalo's milk; three samples were confirmed to be pure cow's milk while the remaining eight samples were mixed cow and buffalo milk. In conclusion, the PCR assays evaluated in this study can be useful for milk inspection to detect cow's milk in water buffalo milk with a detection limit of 0.5%. Also, analysis of market milk samples revealed that adulteration of buffalo milk by mixing with cow's milk or even substituti...
The occurrence of bovine herpes virus-1 (BHV-1) in camels was studied. A total of 186 pneumonic c... more The occurrence of bovine herpes virus-1 (BHV-1) in camels was studied. A total of 186 pneumonic camel lungs were collected from slaughter houses at four different areas in Sudan during 2000-2006. Using sandwich ELISA 1.6% of 186 tested lungs were found positive for BHV-1 antigen, all were from Tambool at Central Sudan. Direct fluorescent antibody test (FAT) was used to confirm the BHV-1 ELISA positives, all ELISA positives were also positive. PCR was used to detect BHV-1 genome with three positive results. BHV-1 was isolated from two camel lungs in MDBK cells. Isolates were identified using ELISA and FAT. Indirect ELISA was used to detect antibodies to BHV-1 in 260 camel sera; 76.9% were found positive. Highest prevalence was observed in sera from Kordofan (84%) then Blue Nile (80%) and Tambool (76.3%). This is the first report for the detection of BHV-1 antigen, genome using PCR, isolation in cell culture and antibodies in camels in Sudan.
This study was undertaken to investigate the role of parainfluenza virus 3 (PIV3) in respiratory ... more This study was undertaken to investigate the role of parainfluenza virus 3 (PIV3) in respiratory infection of camels. A total of 273 lung specimens from camels with pneumonia lesions were collected from slaughterhouses in four different areas of Sudan. In addition, eight specimens were collected from outbreaks of respiratory infection in camels. Using antigen detection sandwich ELISA kits, six out of the 281 specimens tested were positive for the PIV3 antigen (2.1%); the highest prevalence was noted in Eastern Sudan (4.2%), then in Central and Northern Sudan (1.4%). The direct immunofluorescent test (FAT) was used to confirm the positive reactions for PIV3 by ELISA. The polymerase chain reaction (RT-PCR) was applied for the detection of the PIV3 genome in lungs of camels; two out of four samples which were positive by the PIV3 ELISA were also positive by RT-PCR. Virus isolation was attempted for PIV3 in MDBK cells; four specimens yielded cytopathic virus when inoculated onto the cell culture. The cytopathic effect (CPE) consisted of cell rounding, multinucleated cells, sloughing and elongation of cells, and some syncytia were observed on the 3rd to 7th day post-inoculation. Using commercially available indirect ELISA kits for antibodies to PIV3, 495 camel sera were tested, and the seroprevalence detected was 82.2%. The highest seroprevalence was observed in Central (92.6%), then in Eastern (92.2%) and Central to South Sudan (82.5%); the lowest prevalence was found in Northern Sudan (64.8%).
An evaluation of the polymerase chain reaction (PCR) for detection of Brucella melitensis DNA in ... more An evaluation of the polymerase chain reaction (PCR) for detection of Brucella melitensis DNA in bovine and ovine semen was performed. Since semen contains different components that inhibit PCR ampli®cation, a protocol was used to purify Brucella-DNA from bovine and ovine semen samples prior to conducting ampli®cation of the targeted DNA. When separated fractions of naturally Brucella contaminated semen were analyzed by the PCR, most of B. melitensis DNA were present in the seminal¯uid and non-sperm fractions. The PCR examination results for detection of B. melitensis DNA in different semen fractions were compared with the results for traditional cultural methods of Brucella from semen. The PCR was more sensitive than the traditional cultural methods since it detected Brucella-DNA in 12 (10%) out of 120 semen samples while direct culture detected only 7 (5.8%) in the same semen samples. The limit of detection by PCR was 100 CFU/ml of semen. In addition, the results of PCR were available in one day, whereas isolation and identi®cation of Brucella organisms required days or even weeks. The PCR may be used as a supplementary test for detection of B. melitensis in semen.
This study aimed to investigate the occurrence of respiratory syncytial virus (RSV) infections in... more This study aimed to investigate the occurrence of respiratory syncytial virus (RSV) infections in camels in Sudan. A total of 272 camel lung specimens showing pneumonia were collected from slaughter houses at four different areas in Sudan, additionally 8 specimens were collected from outbreaks of respiratory infection in camels. Using sandwich ELISA kits for RSV antigen detection 4 out of 280 tested lungs (1.4%) were positive, all were from Central Sudan (Tambool slaughter house). FAT was used to confirm the ELISA positives. Polymerase chain reaction RT/PCR was applied for the detection of RSV genome in camel lungs; 1 out of 4 ELISA positives was positive by RT/PCR. Using indirect ELISA kits 135 out of 495 (27.3%) camel sera showed antibodies to RSV, highest prevalence was observed in Western (33.5%) then Central (31.6%) and Eastern Sudan (23.5%). Based on the manufacturer specified calculations for OD readings, most of positive sera (90/135) were low reactive (1+). This is the first report for the detection of RSV antigen, genome and antibody in camels in Sudan.
Trypanosoma evansi is enzootic in camels in Egypt, and water buffaloes act as a reservoir for cam... more Trypanosoma evansi is enzootic in camels in Egypt, and water buffaloes act as a reservoir for camel infection. Molecular techniques have contributed towards understanding the epidemiology of T. evansi. Trypanosoma evansi was detected in acute and chronic stages of the disease in male and female mice by polymerase chain reaction (PCR) using two primers. Two experiments were conducted. In experiment I, two groups consisting of 26 female and 26 male mice received 104 trypanosome by I/P inoculation for each mouse. In experiment II, 42 female and 42 male mice were inoculated I/P with 102 trypanosome/mouse. In addition, five mice were kept as uninfected control for each group. Mice were monitored daily for parasitaemia level during the pre-patent period using the micro-haematocrit centrifugation technique (MHCT) and conventional PCR. The primer pairs, (Trypanosoma brucei) TBR1/2 and TeRoTat1.2 (T. evansi Rode Trypanozoon antigen type [RoTat] 1.2), detected the infection after 24 hours ear...
INTRODUCTION High economic lose was resulting due to the reproductive disorders in cattle and buf... more INTRODUCTION High economic lose was resulting due to the reproductive disorders in cattle and buffaloes such as abortions, repeat breeding and infertility (Yoo et al., 2010). Bovine abortion may be due to many causes, but infectious causes are the most important one (Silva et al., 2009). Worldwide, Brucellosis, Leptospirosis, Mycoplasmosis and Listeriosis are considered serious zoonosis and represent an important cause of reproductive losses in animals, especially in Mediterranean countries and Egypt (Gwida et al., 2016). These diseases are characterized by fast spreading, difficulty of control and prevention, time consuming and the cost of treatment (Selim et al., 2014). As a replacement of time consuming traditional methods, PCR has been used as an important diagnostic tool of abortion in cattle (Anderson, 2007). Several single PCR assays have been used for detection of abortion microbial agents, including Brucella species, pathogenic Leptospira species, Mycoplasma species and Listeria monocytogenes (
THE EGYPTIAN JOURNAL OF EXPERIMENTAL BIOLOGY (Zoology)
Graphene oxide Nano-sheets (GOs) have a wide range of industrial, biochemical and medical applica... more Graphene oxide Nano-sheets (GOs) have a wide range of industrial, biochemical and medical applications regarding their unique physical, chemical and biocompatibility properties. For assessment of toxicological potential of graphene oxide nanosheets ninety male mice were divided into six groups; the control and five treated groups: animals of the control group received an intraperitoneal (i.p) injection of 0.2 ml saline solution (0.9% NaCl) once weekly for eight weeks. The treated groups were received an i.p injection of 10, 50, 100, 250, and 500 µg GOs/kg body weight once weekly, for eight weeks. Animals of all groups were sacrificed after 7, 28, and 56-days post treatment. The present study was conducted to evaluate the genotoxic effects of GOs on mice liver cells using alkali comet assay. In addition, physiological investigations and description of hepatic histopathological alternations were carried out. Results revealed that GOs induced DNA damage (DNA fragmentation), represented in dose and time-dependent increase in % tail DNA migration, tail moment and comet tail length. As well as, a diminish in % head DNA in nuclei of liver of GOs-treated mice versus control groups. The effect of GOs on hepatic superoxide dismutase (SOD), catalase (CAT) activities and glutathione (GSH) level indicated slight decrease at low doses (10 and 50 µg) at all time intervals of experimental period as compared to the control groups. The administration of high doses of GOs (100, 250, and 500 µg) indicated a significant diminish in SOD, CAT activities and decrease in GSH level; with subsequent elevation in malondialdehyde (MDA) levels in liver as compared to control groups. Various hepatic histopathological alternations were dose and time dependent in GO-treated mice versus control. In conclusion, the induction of DNA fragmentation in liver denoting the genotoxicity of GOs in dose and time dependent manner. GOs had the ability for inducing various hepatic alternations in a dose and time dependent manner indicating cytotoxicity presumably by oxidative stress. Thus, GOs should be used under strict control and these serious side effects should be taken into consideration when used in vivo treatments.
Experimental and toxicologic pathology : official journal of the Gesellschaft fur Toxikologische Pathologie, Jan 5, 2017
Graphene and graphene-related materials have broadly applied in biomedical purposes due to their ... more Graphene and graphene-related materials have broadly applied in biomedical purposes due to their unique properties, thus safety evaluation of them is crucial. This study was performed to explore the genotoxic and pulmonary toxic potential of different doses of graphene oxide nanosheets' (GOs) in mice.A total of 90 male mature mice were randomly divided into six groups of fifteen mice per each, five groups were intraperitoneally injected by GO at doses of 10, 50, 100, 250 and 500μg/kg b.w once weekly in addition to the control group that was injected intraperitoneally with 0.2ml saline solution. Five animals from each group were euthanized after 7, 28 and 56days post treatment. Evaluation of genotoxicity was performed through detection of chromosomal aberrations in bone marrow while assessment of lung injury was made by determination of DNA fragmentation in lung specimens using the alkali Comet assay, pulmonary oxidative markers estimation and finally histopathological investigat...
The European journal of contraception & reproductive health care : the official journal of the European Society of Contraception, Jan 20, 2014
Objective To study the association between Porphyromonas gingivalis (P. gingivalis) infection and... more Objective To study the association between Porphyromonas gingivalis (P. gingivalis) infection and recurrent miscarriage. Methods This case control study included women with early pregnancy failure admitted for surgical evacuation of retained products of conception. Cases (group 1) included 50 women with unexplained recurrent early miscarriage whereas the control group (group 2) consisted of 50 women with no such history. The evacuated products of conception, subgingival plaques, cervicovaginal secretions and saliva of all participants were examined to detect P. gingivalis deoxyribonucleic acid (DNA) using a polymerase chain reaction. Results The prevalence of P. gingivalis DNA in the chorionic villous tissue samples of group 1 was significantly higher than in group 2 (8 [16%] vs. 1 [2%], respectively; p = 0.036, odds ratio [OR]: 9.3, 95% confidence interval [CI]: 1.1-76.9). The prevalence of P. gingivalis DNA was significantly higher in cervicovaginal secretions of group 1 than in g...
Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis) is the causative agent of p... more Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis) is the causative agent of paratuberculosis, or Johne's disease, in cattle, with potential involvement in cases of Crohn's disease in humans. Johne's disease is found worldwide and is economically important for both beef and dairy industries. In an effort to characterize this important infection in Egypt, we analysed the ecological and genomic features of recent isolates of M. paratuberculosis. In this report, we examined 26 Holstein dairy herds distributed throughout Egypt, from 2010 to 2013. Using PCR analysis of faecal samples, we estimated a mean herd-level prevalence of 65.4 %, with animallevel infection that reached a mean of 13.6 % among animals suffering from diarrhoea. Whole genome sequencing of field isolates identified numerous single nucleotide polymorphisms among field isolates relative to the standard M. paratuberculosis K10 genome. Interestingly, the virulence of M. paratuberculosis isolates from Egypt revealed diverse virulence phenotypes in the murine model of paratuberculosis, with significant differences in tissue colonization, particularly during the chronic stage of infection. Overall, our analysis confirmed that Johne's disease is a newly identified problem in Egypt and indicated that M. paratuberculosis has potentially diverse genotypes that impact its virulence. Further ecological mapping and genomic analysis of M. paratuberculosis will enhance our understanding of the transmission and evolutionary dynamics of this pathogen under natural field conditions.
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Papers by Adel Amin