Diagnosis of Pebrine Disease in Silkworm Using Molecular Methods.

Since pebrine disease, as the most important and dangerous disease in silkworms, spreads horizontally through the spores and vertically through the eggs, combating the disease and eliminating it completely from livestock production has been associated with numerous problems. This project aimed to identify the molecular cause of pebrine disease in silkworms using a sensitive, specific, and accurate method. To this purpose, a 136 bp fragment was selected based on the Nosema bombycis partial SSU rDNA sequence, and a pair of primers was designed. Afterward, using the conventional polymerase chain reaction (PCR) method, the target fragment was amplified and sequenced. After that, to determine the detection sensitivity, using the Real-Time PCR method, 5-fold serial dilutions of N. bombycis DNA were prepared, and the last dilution that produced a fluorescent signal was considered the minimum detection limit. All tests were performed in duplicates. Based on the results of the sensitivity test, the standard curve including Ct values ​​and DNA concentration was used for analysis. Moreover, 80 unknown samples examined by light microscope were evaluated using conventional PCR and Real-Time PCR. Both PCR results showed no amplification for the negative control samples. The findings demonstrated that the lowest detection limit for N. bombycis was less than 6 pg of DNA, while, this amount was 8 ng for conventional PCR. Out of 80 samples examined, 55, 60, and 62 samples were positive for light microscope, conventional PCR, and Real-Time PCR methods, respectively. The findings suggested that the Real-Time PCR method had a higher ability to detect the causative agent of pebrine disease than the conventional PCR method, and both methods were superior to light microscopy. Therefore, due to the fewer steps and higher accuracy of Real-Time PCR, it can be introduced as a suitable method for diagnosing pebrine disease.

Prevalence Determination of m. Hominis and m. Genitalium in the Semen Samples in the Northeast of Iran Using Culture and Multiplex Polymerase Chain Reaction.

Infertility has recently become a growing social and economic world problem. Genital mycoplasmas, such as Mycoplasma hominis and M. genitalium, are most frequently associated with several adverse effects on men’s fertility. The present study aimed to determine the prevalence of M. hominis and M. genitalium in the semen samples in thenortheast of Iran. During thiscross-sectional study from February to May, 2018, 100 semen samples were collected from 100 infertile men in Mashhad, Khorasan Razavi province, northeast of Iran. The presence of M. hominis and M. genitalium was detected by cultivation, polymerase chain reaction (PCR), and Multiplex PCR assays. The colony of mycoplasma was confirmed by Diene’s stain; moreover, arginine hydrolysis, glucose, and urea utilization were evaluated. The following semen indices were analyzed according to World Health Organization guidelines for semen analysis: color, volume, appearance, liquefaction, viscosity, concentration, pH, leukocyte concentration, progressive motility, morphological normality, motile sperm concentration, functional sperm concentration, sperm motility index, and functional sperm. The gene of 16SrRNA (GPO1& MGSO primers) was used as the target gene of the Mycoplasma genus in PCR assay. Multiplex-PCR was performed with a specific primer for conserved regions in the 16SrRNA gene for M. hominis (RNAH1& RNAH2 primers) and the 140-kDa Adhesion Protein Gene for M. genitalium (MG1 & MG2 primers).According to the results,9 (9%) samples were PCR-positive for Mycoplasma spp , while there were 7 (7%) cases isolated by cultivation. M. hominis was detected in 8 (8%) samples by Multiplex PCR, while there was no evidence for M. genitalium. The mean age scores of all infertile and infected men were obtained at 31 and 30 years, respectively. The study could not show any statistical correlation between mycoplasma infection and abnormal semen parameters. The heterogeneity of mycoplasma prevalence in the reports can be ascribed to differences in geographic areas, the sensitivity of the identification method, condition of the group (fertile/infertile), sample size, and operator proficiency. Various results have been reported in numerous studies conducted on the relationship between mycoplasma infection and abnormal semen parameters.

Prevalence of American foulbrood in asymptomatic apiaries of Kurdistan, Iran.

AIM: Paenibacillus larvae subsp. larvae is the etiological agent of American foulbrood (AFB), the most virulent bacterial disease of honey bee brood worldwide. In many countries, AFB is a notifiable disease since it is highly contagious, in most cases incurable, and able to kill affected colonies. The aim of this study was to determine the prevalence of P. larvae subsp. larvae in Kurdistan province apiaries by polymerase chain reaction (PCR) technique. MATERIALS AND METHODS: A total of 100 samples were randomly purchased from apiaries in Kurdistan, Iran. Apiaries were randomly sampled in accordance with the instructions of the veterinary organization from different provinces and were tested using PCR method and an exclusive primer of 16S rRNA for the presence of P. larvae subsp. larvae. RESULTS: The results of this study indicated a low level of contamination with P. larvae subsp. larvae in the Kurdistan province. The number of positive samples obtained by PCR was 2%. CONCLUSION: Therefore, monitoring programs for this honeybee disease in Kurdistan should be developed and implemented to ensure that it is detected early and managed.

Platelet count and neonatal sepsis: a high prevalence of Enterobacter spp.

INTRODUCTION: Sepsis is a common complication in the neonatal intensive care unit. It is most common in the smallest and most premature infants in whom the clinical presentation can be subtle and nonspecific. The objectives of the present study were to identify the most common organisms causing sepsis and their associations with thrombocytopenia. METHODS: This is a retrospective case analysis of blood culture positive patients between March 2003 and July 2007 in a single centre. We enrolled 53 eligible neonates whose blood culture yielded positively for any organism. Blood for the culture was obtained from a peripheral vessel. The data was analysed for differences in platelet and neutrophil count in terms of the microorganisms causing sepsis using chi-square and Fisher's exact tests, analysis of variance and Kruskal-Wallis, as appropriate. RESULTS: The most common organism in the blood culture was Enterobacter spp. with 21 cases (39.6 percent) and the least common was coagulase-positive Staphylococcus spp. The most common organisms in infants with normal weight and early onset sepsis were coagulase-positive Staphylococcus spp. (50 percent and 36.7 percent, respectively), while in other neonates with low birth weight, very low birth weight and late onset sepsis, the most common organism was Enterobacter spp. (40.9 percent, 71.4 percent and 47.8 percent, respectively). The patients with Enterobacter spp. sepsis had a higher incidence of thrombocytopenia. The mortality rate was 15.1 percent (8/53 cases), which was significantly higher among those with the Enterobacter spp. sepsis (five cases, p-value is 0.033). CONCLUSION: Our study shows the changes in the pattern of late onset neonatal infections in the neonatal intensive care unit. Enterobacter spp. is the most common organism causing neonatal sepsis accompanying thrombocytopenia.

Outcome of living kidney transplant: pediatric in comparison to adults.

BACKGROUND: Renal transplantation is the most optimal way to manage children with end-stage renal disease. Despite its benefits, pediatric renal transplantation is a challenge for several transplantation centers in terms of achieving a satisfactory outcome. We sought to compare the long-term outcome of pediatric versus adult recipients who underwent renal transplantation. METHOD: We examined, 2631 recipients of a first kidney from a living donor between 1982 and 2002. The two groups were matched for immunosuppressive therapy and number of HLA mismatches. The patients were divided into a pediatric (n=301; age 18 years) to compare 5-year patient and graft survivals. RESULTS: The mean ages of the pediatric and adult groups were 40 +/- 13 and 14 +/- 13 years, respectively. The 5-year graft survival was lower among the pediatric versus the adult group (56% vs 68%; P=.015) with no difference in patient survival (88% vs 86%; P>.05). CONCLUSION: The poorer graft survival in pediatric transplantation may be due to the nature of pediatric transplantation, in terms of inconsistent adherence to medication regimens, worse side effects of medications, higher rate of graft rejection due to recurrent disease, and more intense immunoreactivity of children.