The first isolation and detection of Ornithobacterium rhinotracheale from swollen head syndrome-infected broiler flocks in Iraq

Background and Aim: The swollen head syndrome (SHS) makes up complex diseases that infect the upper respiratory tract in poultry and causes several economic losses. Furthermore, this syndrome is considered one of the multifactorial etiological agents. Therefore, this study isolated and molecularly detected Ornithobacterium rhinotracheale (ORT) in poultry. Materials and Methods: This study was conducted at 67 broiler farms that had birds observed to be infected with the SHS from September 2018 until August 2019. Subsequently, swabs were collected from their trachea, infraorbital sinuses, and lungs, after which obtained samples were treated through two methods: (a) The direct method, by uploading samples on FTA cards, and the indirect method using a transport media. Afterward, reverse transcription-polymerase chain reaction (RT-PCR) was used to analyze the directly treated samples; howeverAQ1, the culture method, followed by PCR, was used to analyze the indirectly treated samples. Next, a partial 16S RNA gene was isolated using four positive PCR products, after which the effect of 16 antibiotics was studied on the seven local ORT strains isolated. Results: The quantity of ORT isolated using the direct method was 28 (41.7%) samples, which were all positive for the strain. Identification was by direct molecular identification (RT-PCR) from samples loaded on FTA cards. Alternatively, 7 (10.4%) ORTs were detected from the indirect method, as obtained using the culture method and biochemical tests. Then, PCR was subsequently used to confirm the results. As observed, 784 bp bands were shown for all seven ORT isolates. Furthermore, results revealed a significant difference in the detection of ORT strains between direct and indirect methods, with p-value (<0.05) and standard deviation of the error ±0.038 for the direct, then ±0.061 for the indirect method. For further analysis on the strain types, four 784 bp PCR products were taken, then partial 16S ribosomal sequence typing was conducted. All these four strains were found to be recorded in NCBI for the 1st time as a local Iraqi strain, with accession numbers (MN931657, MN931656, MN931655, and MN931654). Notably, results also showed that all isolated strains were multidrug-resistant. Conclusion: From the results, ORT is proposed to be implicated as one of the etiological factors that cause SHSs in poultry. Phylogenetic analysis of the current ORT bacterial strains also showed that they are closely related to the Egyptian isolates.

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PROTEIN “SOLUBILIZATION” AND CALCIUM AND MAGNESIUM PRECIPITATION BY “CHLORINATED CLEANERS”1

Journal of Milk and Food Technology ◽

10.4315/0022-2747-30.10.310 ◽

1967 ◽

Vol 30 (10) ◽

pp. 310-316 ◽

Cited By ~ 2

Author(s):

E. O. Wright ◽

W. S. LaGrange ◽

Connie Dennis ◽

E. W. Bird ◽

D. H. Hotchkiss

Keyword(s):

Sodium Hypochlorite ◽

Indirect Method ◽

Direct Method ◽

Hard Water ◽

Magnesium Content ◽

The Other ◽

Protein Solubilization ◽

Significant Difference ◽

Calcium And Magnesium

Summary It has been stated that chlorinated cleansers increase the “protein solubilization” properties of cleanser solutions to which they are added (7). Chlorinated cleansers, however, are alkaline materials which increase the active alkalinity of the solutions to which they are added; this seems to have been overlooked. The study reported here was undertaken to determine whether the added available chlorine or the increase in active alkalinity was responsible for the increase in protein solubilization. Two chlorine sources were employed: Sodium hypochlorite (Chlorox) and chlorinated TSP. The approximate amounts by which these chlorine sources increased the active alkalinity (as % NaOH) at the 100 ppm and 200 ppm levels were: Chlorox, 0.0091% and 0.0199%, and chlorinated TSP, 0.0425% and 0.0800%. When the active alkalinity was the same in two solutions , there was no significant difference in the amounts of protein they would dissolve, even though one of the solutions contained available chlorine and the other did not. However, the pairs of solutions containing the same percentage of active alkalinity as the solution containing 200 ppm available chlorine solubilized significantly more protein (p <0.06) than did the pairs which contained the same active alkalinity as the solution containing 100 ppm; this is attributed to the higher active alkalinity of the former. These results were the same whether Chlorox or chlorinated TSP was the chlorine source, whether the direct or indirect method of determining protein solubilization was employed, and in the direct method, whether skim or homogenized milk was the soiling agent. On the basis of these data, the increase in active alkalinity, not the added available chlorine appears responsible for the increase in protein solubilization when a chlorine source is added to an alkaline cleanser. No calcium remained in any of the spent cleansers from the direct method of evaluating protein solubilization. In general, the amount of magnesium remaining decreased as the active alkalinity of the solution increased. There was no significant difference in magnesium content between the chlorine-containing and chlorine-free solutions having the same active alkalinity. Chlorine sources apparently minimize precipitation of hard water cations onto equipment surfaces.

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Linkage between H. pylori Infection and TNF-α polymorphism in The Pregnant Women

International Journal of Research in Pharmaceutical Sciences ◽

10.26452/ijrps.v9ispl1.1298 ◽

2018 ◽

Vol 9 (SPL1) ◽

Author(s):

Ghaidaa Raheem Lateef ◽

Azhar Omaran Al-Thahab

Keyword(s):

Pregnant Women ◽

Serum Antibody ◽

Rapid Test ◽

Negative Group ◽

Positive Group ◽

Gynecology And Obstetrics ◽

Tnf Α ◽

Significant Difference ◽

Pcr Products ◽

H Pylori

A study was performed on 100 pregnant women in the outpatient department of gynecology and obstetrics of Maternity and Children Hospital in Al-Diwaniya City during the period between (March to September 2016). One hundred blood samples (50 for patients and 50 for control) were collected under the supervision of the treating gynecologist. The detection of Helicobacter. pylori was done by the use of the serum antibody Rapid test. The results showed that 50 (100%) were positive and 50 (100%) were negative for H. pylori in above method.All blood of patients and control samples were used for the extraction of genomic DNA,where the 107 bp PCR product size. Genotyping of the TNF-α-308 SNP (G/A)was performed by restriction fragment length polymorphism PCR (RFLP-PCR). PCR products were digested with restr NcoI iction enzyme. Individuals with the TNF-α-308(GG) homozygote produced digested DNA bands at 80,and 20 bp bp. A heterozygous genotype ofTNF-α-308 (GA)produced 107 bp,80 bp,and 20 bp bands. Individuals with the TNF-α-308 (AA) homozygote genotype had no amplicon digested and generated only one band of 107 bp. There was a significant difference in the frequency of the TNF-α-308(GG)genotype between H. pylori positive group and H. pylori negative group(72%,78% respectively). Also for GA genotype,there was a significant difference between H. pylori positive group and H. pylori negative group(24%,18% respectively). Concerning the frequency of the TNF-α-308 (AA)genotype between H. pylori positive group and H. pylori negative group,there was no significant difference between the two groups.

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MICROSCOPIC AND MOLECULAR DIAGNOSIS OF Ascaridia spp. IN DOMESTIC PIGEONS(Columba livia domestica) IN BAGHDAD CITY,IRAQ

IRAQI JOURNAL OF AGRICULTURAL SCIENCES ◽

10.36103/ijas.v51i4.1101 ◽

2020 ◽

Vol 51 (4) ◽

pp. 1220-1225

Author(s):

Faraj & Al- Amery

Keyword(s):

Molecular Diagnosis ◽

Columba Livia ◽

Ascaridia Galli ◽

Molecular Assay ◽

Infection Rates ◽

Conventional Pcr ◽

Significant Difference ◽

Pcr Products ◽

Males And Females ◽

Domestic Pigeons

Ascaridiosis is a very important parasitic disease of birds, it is caused by Ascaridia. This study was conducted to identify the Ascaridia species by microscopic and molecular assay in Baghdad city. One hundred and sixty fecal samples were collected from domestic pigeons during the period from 1/1/ 2019 to 31/3/ 2019. Results showed that the rate of infection for Ascaridia spp. 15.62% by microscopic examination. Significant difference was observed in infection rates between males and females pigeons. Fifty samples randomly selected and subjected to molecular diagnosis of Ascaridia spp.. Molecular examination results, the total infection rate showed 16%(8/50). The eight positive PCR products were sequenced and deposited in Gene bank data base, phylogenic analysis demonstrated that 4 sequences belongs to Ascaridia galli ( MK918635.1, MK918636.1, MK918847.1, MK919081.1), while 2 (MK919199.1, MK919200.1) belong to Ascaridia nymphii and 2 (MK919207.1, MK919264.1) belong to Ascaridia numidae. It is the first study in Iraq to diagnosis of Ascaridia nymphii and Ascaridia numidae in domesticed pigeons by using conventional PCR.

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Parathyroid Ca2+-conducting currents are modulated by muscarinic receptor agonists and antagonists

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.1997.273.5.e880 ◽

1997 ◽

Vol 273 (5) ◽

pp. E880-E890 ◽

Cited By ~ 1

Author(s):

Wenhan Chang ◽

Tsui-Hua Chen ◽

Stacy A. Pratt ◽

Benedict Yen ◽

Michael Fu ◽

...

Keyword(s):

Muscarinic Receptors ◽

Inositol Phosphate ◽

Receptor Protein ◽

Dependent Manner ◽

Rt Pcr ◽

Pipette Solution ◽

Pcr Products ◽

Inositol Phosphate Accumulation ◽

Receptor Cdna ◽

Dose Dependent

Parathyroid cells express Ca2+-conducting cation currents, which are activated by raising the extracellular Ca2+ concentration ([Ca2+]o) and blocked by dihydropyridines. We found that acetylcholine (ACh) inhibited these currents in a reversible, dose-dependent manner (50% inhibitory concentration ≈10−8 M). The inhibitory effects could be mimicked by the agonist (+)-muscarine. The effects of ACh were blunted by the antagonist atropine and reversed by removing ATP from the pipette solution. (+)-Muscarine enhanced the adenosine 3′,5′-cyclic monophosphate (cAMP) production by 30% but had no effect on inositol phosphate accumulation in parathyroid cells. Oligonucleotide primers, based on sequences of known muscarinic receptors (M1-M5), were used in reverse transcriptase-polymerase chain reaction (RT-PCR) to amplify receptor cDNA from parathyroid poly (A)+ RNA. RT-PCR products displayed >90% nucleotide sequence identity to human M2- and M4-receptor cDNAs. Expression of M2-receptor protein was further confirmed by immunoblotting and immunocytochemistry. Thus parathyroid cells express muscarinic receptors of M2 and possibly M4 subtypes. These receptors may couple to dihydropyridine-sensitive, cation-selective currents through the activation of adenylate cyclase and ATP-dependent pathways in these cells.

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Molecular detection of cattle Sarcocystis spp. in North-West Italy highlights their association with bovine eosinophilic myositis

Parasites & Vectors ◽

10.1186/s13071-021-04722-5 ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

Selene Rubiola ◽

Tiziana Civera ◽

Felice Panebianco ◽

Davide Vercellino ◽

Francesco Chiesa

Keyword(s):

18S Rdna ◽

Inflammatory Myopathy ◽

Molecular Techniques ◽

Diaphragm Muscle ◽

Economic Losses ◽

Intermediate Hosts ◽

Slaughter Cattle ◽

Significant Difference ◽

Pcr Targeting

Abstract Background Cattle are intermediate hosts of six Sarcocystis species, among which Sarcocystis hominis and Sarcocystis heydorni can infect humans through the consumption of raw or undercooked meat. In addition to the zoonotic potential, there is increasing interest in these protozoa because of the evidence supporting the role of Sarcocystis spp. in the occurrence of bovine eosinophilic myositis (BEM), a specific inflammatory myopathy which leads to carcass condemnation and considerable economic losses. Actually, all the prevalence studies carried out on cattle in Italy have been based on either morphological or 18S rDNA-based molecular techniques, most likely leading to misidentification of closely related species. Therefore, there is a strong need for new data on the prevalence of the different Sarcocystis spp. in cattle in Italy and their association with bovine eosinophilic myositis. Methods To reach our aim, individual striated muscle samples from BEM condemned carcasses (N = 54) and diaphragm muscle samples from randomly sampled carcasses (N = 59) were obtained from Northwest Italy slaughterhouses. Genomic DNA was extracted and analyzed by multiplex-PCR targeting 18S rDNA and cox1 genes. PCR products amplified using the genus-specific primer set in absence of the specific fragment for S. hirsuta, S. cruzi, S. hominis or S. bovifelis were sequenced to achieve species identification. Results Sarcocystis DNA was detected in 67.8% of the samples from slaughter cattle and in 90.7% of the samples from BEM condemned carcasses. S. cruzi was identified as the most prevalent species in slaughter cattle (61%), followed by S. bovifelis (10.2%), S. hominis (8.5%) and S. hirsuta (1.7%). Notably, among the different Sarcocystis spp. detected, the presence of S. bovifelis and S. hominis was significantly higher in samples isolated from BEM condemned carcasses (46.3% and 40.7% respectively), while there was no statistically significant difference between the presence of S. cruzi or S. hirsuta in BEM condemned carcasses (42.6% and 1.8%, respectively) and randomly sampled carcasses. Furthermore, DNA sequence analysis revealed the presence of a putative new species in two carcasses. Conclusions Our study contributes to updating the data on the prevalence of the different Sarcocystis spp. in cattle in Italy, highlighting the presence of three Sarcocystis spp., S. cruzi, S. hominis and S. bovifelis, in BEM lesions and allowing us to speculate on the possible role of S. hominis and S. bovifelis as the major sarcosporidian species involved in bovine eosinophilic myositis. Graphic Abstract

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Duplex-RT-PCR assay for the simultaneous detection and discrimination of Brome mosaic virus and Cocksfoot mottle virus in cereal plants

Molecular Biology Reports ◽

10.1007/s11033-021-06485-9 ◽

2021 ◽

Author(s):

Katarzyna Trzmiel

Keyword(s):

Mosaic Virus ◽

Simultaneous Detection ◽

Brome Mosaic Virus ◽

Mottle Virus ◽

Grass Species ◽

Replicase Gene ◽

Rt Pcr ◽

Pcr Products ◽

Cereal Plants ◽

Cocksfoot Mottle Virus

AbstractBrome mosaic virus (BMV) and cocksfoot mottle virus (CfMV) are pathogens of grass species including all economically important cereals. Both viruses have been identified in Poland therefore they create a potential risk to cereal crops. In this study, a duplex—reverse transcription—polymerase chain reaction (duplex-RT-PCR) was developed and optimized for simultaneous detection and differentiation of BMV and CfMV as well as for confirmation of their co-infection. Selected primers CfMVdiag-F/CfMVdiag-R and BMV2-F/BMV2-R amplified 390 bp and 798 bp RT-PCR products within coat protein (CP) region of CfMV and replicase gene of BMV, respectively. Duplex-RT-PCR was successfully applied for the detection of CfMV-P1 and different Polish BMV isolates. Moreover, one sample was found to be co-infected with BMV-ML1 and CfMV-ML1 isolates. The specificity of generated RT-PCR products was verified by sequencing. Duplex-RT-PCR, like conventional RT-PCR, was able to detect two viruses occurring in plant tissues in very low concentration (as low as 4.5 pg/µL of total RNA). In contrast to existing methods, newly developed technique offers a significant time and cost-saving advantage. In conclusion, duplex-RT-PCR is a useful tool which can be implemented by phytosanitary services to rapid detection and differentiation of BMV and CfMV.

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RT-PCR analysis of Deformed wing virus in honeybees (Apis mellifera) and mites (Varroa destructor)

Journal of General Virology ◽

10.1099/vir.0.81401-0 ◽

2005 ◽

Vol 86 (12) ◽

pp. 3419-3424 ◽

Cited By ~ 210

Author(s):

Constanze Yue ◽

Elke Genersch

Keyword(s):

Varroa Destructor ◽

Pcr Analysis ◽

Body Parts ◽

Rt Pcr ◽

Larval Food ◽

Viral Pathogen ◽

Deformed Wing Virus ◽

Significant Difference ◽

Almost All ◽

Viral Sequences

Deformed wing virus (DWV) is a honeybee viral pathogen either persisting as an inapparent infection or resulting in wing deformity. The occurrence of deformity is associated with the transmission of DWV through Varroa destructor during pupal stages. Such infections with DWV add to the pathology of V. destructor and play a major role in colony collapse in the course of varroosis. Using a recently developed RT-PCR protocol for the detection of DWV, individual bees and mites originating from hives differing in Varroa infestation levels and the occurrence of crippled bees were analysed. It was found that 100 % of both crippled and asymptomatic bees were positive for DWV. However, a significant difference in the spatial distribution of DWV between asymptomatic and crippled bees could be demonstrated: when analysing head, thorax and abdomen of crippled bees, all body parts were always strongly positive for viral sequences. In contrast, for asymptomatic bees viral sequences could be detected in RNA extracted from the thorax and/or abdomen but never in RNA extracted from the head. DWV replication was demonstrated in almost all DWV-positive body parts of infected bees. Analysing individual mites for the presence of DWV revealed that the percentage of DWV-positive mites differed between mite populations. In addition, it was demonstrated that DWV was able to replicate in some but not all mites. Interestingly, virus replication in mites was correlated with wing deformity. DWV was also detected in the larval food, implicating that in addition to transmission by V. destructor DWV is also transmitted by feeding.

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A survey for bvdv antibodies in cattle farms in Slovakia and genetic typing of bvdv isolates from imported animals

Acta Veterinaria Hungarica ◽

10.1556/avet.51.2003.2.11 ◽

2003 ◽

Vol 51 (2) ◽

pp. 229-236 ◽

Cited By ~ 8

Author(s):

Š. Vilček ◽

Jana Mojžišová ◽

Viera Bajová ◽

Š. Paulík ◽

L. Strojný ◽

...

Keyword(s):

Bovine Viral Diarrhoea Virus ◽

Computer Assisted ◽

Serological Survey ◽

Bovine Viral Diarrhoea ◽

Rt Pcr ◽

Serum Samples ◽

Genetic Typing ◽

Diarrhoea Virus ◽

Pcr Products ◽

Cattle Farms

A serological survey for bovine viral diarrhoea virus (BVDV) antibodies on a collection of 1295 serum samples obtained from 6-12 months old cattle originating from 45 farms in Slovakia was carried out. On 13 farms more than 90% of the examined animals were seropositive, on 14 farms 71-90% seroprevalence was observed, on 13 farms only 50-70% animals were found to be positive for BVDV antibodies, while the remaining 5 farms showed fewer than 50% seropositive animals. The average incidence of BVDV antibodies (around 70%) was similar as determined 30 years ago. Of 84 serum samples from seronegative animals originating from 14 farms in which 70-98% seropositivity was observed, six were positive in Ag-BVDV ELISA indicating persistently infected (PI) cattle. On a farm to which animals were imported from abroad, a BVD outbreak was observed. Of 110 animals tested, four were positive in Ag-ELISA indicating the presence of PI cattle on this farm. Genetic typing of two isolates from imported animals performed by RT-PCR (324/326 primers from 5´-UTR), sequencing of PCR products and computer-assisted phylogenetic analysis revealed that they belong to BVDV-1h group.

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