Epidemiological Molecular Study of Combined Severe Immune Deficiency in Arabian Horses in Tunisia

2021 ◽  
Author(s):  
Ikram Bensouf ◽  
Hatem Ouled Ahmed ◽  
Atef Malek ◽  
Sarra Torjemane ◽  
Faten Lasfar ◽  
...  
Keyword(s):  
Recessive Gene ◽  
Molecular Study ◽  
Peripheral Blood Lymphocytes ◽  
Base Pairs ◽  
Gene Encoding ◽  
Horse Population ◽  
Knowing That ◽  
Spin Column ◽  
Arabian Horse ◽  
Pcr Products

Background: Severe Combined Immuno Deficiency (SCID) is a heritable deficiency transmitted through autosomal recessive gene, carried on by purebred Arabian and crossbred horses. A deletion of five base pairs at the gene encoding the catalytic subunit of the Protein Kinase DNA-dependent (DNA-PKcs) is responsible for this disease and SCID-affected animals always die in the first six months of life. Considering this problem, it is important to perform a molecular epidemiological study to estimate the frequency of the SCID allele in Arabian purebred horses in Tunisia.Methods: The DNA of the peripheral blood lymphocytes of 164 purebred Arabian horses belonging to the Sidi Thabet stud was extracted by the spin-column method, in order to verify the quality of the DNAs used, the samples were dosed by a spectrophotometer. The amplification of the DNA was carried out by two specific primers in a Polymerase Chain Reaction (PCR) to flank the deletion zone of the DNA sample by adding to each PCR sample a marker or a standard of size knowing that the fragment we wanted to amplify is composed of 163 base pairs. Then, the PCR products were sequenced using an automatic sequencer. Result: By analyzing the electropherograms results, we noted the absence of the SCID deletion in the studied group of Arab purebreds. Hence, it is essential to carry out molecular screening of the SCID deletion in other sites in order to determine its prevalence in the country’s purebred Arabian horse population.

MOLECULAR STUDY OF HUMAN HEAD LICE, PEDICULUS HUMANUS CAPITIS COMPARISON WITH GOAT SUCKING LICE, LINOGNATHUS STENOPSIS (BURMEISTER) STRAINS

2020 ◽  
pp. 132-139
Keyword(s):  
Genetic Difference ◽  
Human Head ◽  
Molecular Study ◽  
Head Lice ◽  
Pediculus Humanus Capitis ◽  
School Students ◽  
Pediculus Humanus ◽  
Sucking Lice ◽  
Pcr Products ◽  
Amplicon Size

In this study, only (122) out of (915) primary school students were shown to be infected with head lice Pediculus. humanus capitis. The number and percentage of infected males were 46 (11.3%), while the number and percentage of infected females were 76 (14.9%). The results in our study also showed that the number and percentage of goats infected with goat sucking lice, Linognathus stenopsis was 70 (21.7%) of the total 322 animals, with the highest number and percentage among female goats 44 (62.9%) compared to the male goats 26 (37.1%). The study demonstrated that the rate of genetic difference between the studied samples was 89% and the similarity rate was 11%. Detection of OP-K01 gene pieces by PCR products showed that the amplicon size was 520 bp for P. humanus capitis isolated from humans, while the detection of OP-E20 and OP-M05 gene pieces with PCR product showed the lowest amplicon size 230 bp for Linognathus stenosis isolated from goats.

scholarly journals Serological Survey of Leptospira Infection in Arabian Horses in Poland

Pathogens ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 688
Author(s):  
Bernard Wasiński ◽  
Katarzyna Paschalis-Trela ◽  
Jan Trela ◽  
Michał Czopowicz ◽  
Jerzy Kita ◽  
...  
Keyword(s):  
Companion Animals ◽  
Laboratory Diagnosis ◽  
Serological Survey ◽  
Serum Samples ◽  
Horse Population ◽  
Zoonotic Infections ◽  
Clinical Presentations ◽  
Arabian Horse ◽  
Breeding Farms

Leptospirosis is one of the most common zoonotic infections worldwide, including in most livestock, some companion animals, horses, wildlife, and humans. Epidemiological estimation of its prevalence in all species is difficult due to the variety of clinical presentations and challenges regarding laboratory diagnosis. The purpose of this study was to measure the seroprevalence of leptospiral infection in Arabian horses kept in the largest breeding farms in Poland, representing over 15% of the Polish Arabian horse population. Leptospira antibodies were detected by MAT (cut-off 1:100) in 33.2% of serum samples (204 of 615 animals) (CI 95%: 29.6–37.0%), most frequently reacting with the serovar Grippotyphosa, similar to previous reports in populations of randomly selected horses. These results indicated high Leptospira seropositivity, thus, although any form of clinical leptospirosis is rare, it may be postulated that the leptospiral exposure is widespread.

Differential expression of the human gonadotropin alpha gene in ectopic and eutopic cells

1985 ◽  
Vol 5 (11) ◽  
pp. 3157-3167
Author(s):  
R B Darnell ◽  
I Boime
Keyword(s):  
Hela Cells ◽  
Cell Types ◽  
Base Pairs ◽  
Gene Encoding ◽  
Basal Expression ◽  
Bacterial Enzyme ◽  
Specific Induction ◽  
Alpha Gene ◽  
Cat Gene ◽  
Jar Cells

We have analyzed the regulation of the alpha gonadotropin gene in eutopic placental cells and ectopic tumor cells by constructing a series of plasmid vectors containing alpha genomic 5' flanking DNA placed upstream of the gene encoding the bacterial enzyme chloramphenicol acetyltransferase (CAT). These plasmid DNAs were transfected into a eutopic (JAr) and an ectopic (HeLa) cell line. Both cell types expressed the CAT gene from plasmid constructs containing as much as 1,500 base pairs (bp) and as little as 140 bp of alpha 5' flanking DNA; JAr cells were considerably more efficient than HeLa cells. Ectopic and eutopic cells differed qualitatively in their expression from these alpha-CAT constructs when cells were treated with cAMP or butyrate. Butyrate induced alpha expression in HeLa cells but not in JAr cells, while cAMP induced expression in JAr cells. These results are consistent with and extend previous observations suggesting that there are cell-specific differences in the regulation of alpha gene expression in ectopic and eutopic cells. However, by using deletion constructs of the alpha-CAT gene, we found that the basal expression and cell-specific induction of the alpha gene in ectopic and eutopic cells were dependent on the same 140 bp of alpha 5' flanking DNA. These 140 bp were sequenced and found to contain a 9-bp stretch of DNA homologous with the consensus viral enhancer sequence. Such features of alpha expression common to both ectopic and eutopic cells may be involved in the coordinate expression of the alpha gene and the tumorigenic phenotype observed in each cell type.

Glucose-regulated protein (GRP94 and GRP78) genes share common regulatory domains and are coordinately regulated by common trans-acting factors

1989 ◽  
Vol 9 (5) ◽  
pp. 2153-2162
Author(s):  
S C Chang ◽  
A E Erwin ◽  
A S Lee
Keyword(s):  
Gene Promoters ◽  
Base Pairs ◽  
Gene Encoding ◽  
Regulatory Domains ◽  
Coordinated Expression ◽  
Common Domain ◽  
Glucose Regulated Protein ◽  
Competition Assays

We isolated the promoter of the human gene encoding the 94,000-dalton glucose-regulated protein (GRP94). The 5'-flanking region important for its expression was identified by deletion analysis. Comparison of the promoters of the genes for GRP78 and GRP94 derived from human, rat, and chicken cells revealed a common domain of 28 base pairs within the putative regulatory regions of both genes. This domain has been shown to interact with protein factors in the promoter of the gene for GRP78. Since the genes for GRP94 and GRP78 are transcriptionally regulated with similar kinetics under a variety of stress conditions, we are interested in examining the possible mechanisms for their coordinated expression. Through in vitro and in vivo competition assays, we found that the protein factors which interact with the promoter of the gene for GRP94 also have affinity for the conserved domain of the promoter of the gene for GRP78. These findings suggest that the genes for GRP94 and GRP78 are coordinately regulated through common trans-acting factors which recognize a common regulatory domain of glucose-regulated protein gene promoters.

scholarly journals Inequality in the Frequency of the Open States Occurrence Depends on Single 2H/1H Replacement in DNA

Molecules ◽  
2020 ◽  
Vol 25 (16) ◽  
pp. 3753 ◽  
Author(s):  
Alexander Basov ◽  
Mikhail Drobotenko ◽  
Alexandr Svidlov ◽  
Eugeny Gerasimenko ◽  
Vadim Malyshko ◽  
...  
Keyword(s):  
Mathematical Modeling ◽  
False Negative ◽  
Deuterium Atom ◽  
Base Pairs ◽  
Gene Encoding ◽  
Negative Results ◽  
False Negative Results ◽  
Nitrogenous Base ◽  
Open States ◽  
Different Parts

In the present study, the effect of 2H/1H isotopic exchange in hydrogen bonds between nitrogenous base pairs on occurrence and open states zones dynamics is investigated. These processes are studied using mathematical modeling, taking into account the number of open states between base pairs. The calculations of the probability of occurrence of open states in different parts of the gene were done depending on the localization of the deuterium atom. The mathematical modeling study demonstrated significant inequality (dependent on single 2H/1H replacement in DNA) among three parts of the gene similar in length of the frequency of occurrence of the open states. In this paper, the new convenient approach of the analysis of the abnormal frequency of open states in different parts of the gene encoding interferon alpha 17 was presented, which took into account both rising and decreasing of them that allowed to make a prediction of the functional instability of the specific DNA regions. One advantage of the new algorithm is diminishing the number of both false positive and false negative results in data filtered by this approach compared to the pure fractile methods, such as deciles or quartiles.

scholarly journals Molecular Analysis of Ammonia-Oxidizing Bacteria of the β Subdivision of the Class Proteobacteria in Compost and Composted Materials

1999 ◽  
Vol 65 (2) ◽  
pp. 396-403 ◽  
Author(s):  
George A. Kowalchuk ◽  
Zinaida S. Naoumenko ◽  
Piet J. L. Derikx ◽  
Andreas Felske ◽  
John R. Stephen ◽  
...  
Keyword(s):  
Denaturing Gradient Gel Electrophoresis ◽  
Ammonia Oxidizer ◽  
Ammonia Oxidizing Bacteria ◽  
Nitrogen Transformations ◽  
Rt Pcr ◽  
Gene Encoding ◽  
Specific Pcr ◽  
Pcr Products ◽  
Gradient Gel Electrophoresis ◽  
Almost All

ABSTRACT Although the practice of composting animal wastes for use as biofertilizers has increased in recent years, little is known about the microorganisms responsible for the nitrogen transformations which occur in compost and during the composting process. Ammonia is the principle available nitrogenous compound in composting material, and the conversion of this compound to nitrite in the environment by chemolithotrophic ammonia-oxidizing bacteria is an essential step in nitrogen cycling. Therefore, the distribution of ammonia-oxidizing members of the β subdivision of the class Proteobacteriain a variety of composting materials was assessed by amplifying 16S ribosomal DNA (rDNA) and 16S rRNA by PCR and reverse transcriptase PCR (RT-PCR), respectively. The PCR and RT-PCR products were separated by denaturing gradient gel electrophoresis (DGGE) and were identified by hybridization with a hierarchical set of oligonucleotide probes designed to detect ammonia oxidizer-like sequence clusters in the genera Nitrosospira and Nitrosomonas. Ammonia oxidizer-like 16S rDNA was detected in almost all of the materials tested, including industrial and experimental composts, manure, and commercial biofertilizers. A comparison of the DGGE and hybridization results after specific PCR and RT-PCR suggested that not all of the different ammonia oxidizer groups detected in compost are equally active. amoA, the gene encoding the active-site-containing subunit of ammonia monooxygenase, was also targeted by PCR, and template concentrations were estimated by competitive PCR. Detection of ammonia-oxidizing bacteria in the composts tested suggested that such materials may not be biologically inert with respect to nitrification and that the fate of nitrogen during composting and compost storage may be affected by the presence of these organisms.

scholarly journals Detection and Differentiation of Primate α-herpesviruses by PCR

1997 ◽  
Vol 9 (3) ◽  
pp. 225-231 ◽  
Author(s):  
Daria H. Black ◽  
R. Eberle
Keyword(s):  
Pcr Primers ◽  
Restriction Pattern ◽  
Restriction Enzyme Digestion ◽  
Gene Encoding ◽  
B Virus ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Unique Restriction ◽  
Polymerase Chain ◽  
Simplex Virus

A rapid method for detection and differentiation of 5 primate αpha-herpesviruses (human herpes simplex virus types 1 and 2 [HSV1, HSV2], green monkey simian agent 8, baboon herpesvirus 2 [HVP2], and macaque B virus [BV]) was developed utilizing the polymerase chain reaction (PCR). PCR primers were located in conserved regions of the gene encoding the glycoprotein B, which flanks an intervening region that is highly divergent among the 5 viruses. Amplified PCR products from the 5 viruses were readily differentiated by their unique restriction enzyme digestion patterns. No variation in digestion patterns was noted among strains of HSV1, HSV2, or HVP2. One clinical isolate of BV exhibited variation in a single restriction site, but its overall restriction pattern remained typical of BV. This method (PCR/RFLP) allowed the presence of herpesvirus DNA in clinical swabs from primates to be readily detected and the virus unambiguously identified.

scholarly journals GadY, a Small-RNA Regulator of Acid Response Genes in Escherichia coli

2004 ◽  
Vol 186 (20) ◽  
pp. 6698-6705 ◽  
Author(s):  
Jason A. Opdyke ◽  
Ju-Gyeong Kang ◽  
Gisela Storz
Keyword(s):  
Escherichia Coli ◽  
Stationary Phase ◽  
Small Rna ◽  
Acid Resistance ◽  
Base Pairs ◽  
Gene Encoding ◽  
Mrna Accumulation ◽  
Long Form ◽  
Genes Encoding ◽  
Opposite Strand

ABSTRACT A previous bioinformatics-based search for small RNAs in Escherichia coli identified a novel RNA named IS183. The gene encoding this small RNA is located between and on the opposite strand of genes encoding two transcriptional regulators of the acid response, gadX (yhiX) and gadW (yhiW). Given that IS183 is encoded in the gad gene cluster and because of its role in regulating acid response genes reported here, this RNA has been renamed GadY. We show that GadY exists in three forms, a long form consisting of 105 nucleotides and two processed forms, consisting of 90 and 59 nucleotides. The expression of this small RNA is highly induced during stationary phase in a manner that is dependent on the alternative sigma factor σS. Overexpression of the three GadY RNA forms resulted in increased levels of the mRNA encoding the GadX transcriptional activator, which in turn caused increased levels of the GadA and GadB glutamate decarboxylases. A promoter mutation which abolished gadY expression resulted in a reduction in the amount of gadX mRNA during stationary phase. The gadY gene was shown to overlap the 3′ end of the gadX gene, and this overlap region was found to be necessary for the GadY-dependent accumulation of gadX mRNA. We suggest that during stationary phase, GadY forms base pairs with the 3′-untranslated region of the gadX mRNA and confers increased stability, allowing for gadX mRNA accumulation and the increased expression of downstream acid resistance genes.

Molecular biology can differentiate morphologically indistinguishable myxosporean species: Myxobolus elegans and M. hungaricus (Short communication)

2002 ◽  
Vol 50 (1) ◽  
pp. 59-62 ◽  
Author(s):  
Edit Eszterbauer
Keyword(s):  
Ribosomal Rna ◽  
Dna Sequences ◽  
Valid Species ◽  
Base Pairs ◽  
The Family ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Biological Methods ◽  
Myxosporean Species

Two, morphologically indistinguishable myxosporean species, Myxobolus elegans Kashkovsky, 1966 and M. hungaricus Jaczó, 1940 were differentiated using molecular biological methods. Polymerase chain reaction (PCR) with primers specific for the family Myxobolidae was used to amplify an approximately 1600 base pairs (bp) long fragment of the 18S ribosomal RNA gene. In restriction fragment length polymorphism (RFLP) study with HinfI, MspI and TaqI enzymes, the two parasite species were easily distinguishable. The genetic distinctness was also confirmed by the DNA sequence of their PCR products. Although M. elegans and M. hungaricus are morphologically very similar, based on the results of the PCR-RFLP and the DNA sequences, we concluded that they are valid species.

scholarly journals Random sampling of the Central European bat fauna reveals the existence of numerous hitherto unknown adenoviruses

2015 ◽  
Vol 63 (4) ◽  
pp. 508-525 ◽  
Author(s):  
Márton Z. Vidovszky ◽  
Claudia Kohl ◽  
Sándor Boldogh ◽  
Tamás Görföl ◽  
Gudrun Wibbelt ◽  
...  
Keyword(s):  
Rectal Swab ◽  
Polymerase Gene ◽  
Internal Organs ◽  
Gene Encoding ◽  
Closely Related Species ◽  
Central European ◽  
Extant Species ◽  
High Prevalence ◽  
Pcr Products ◽  
And Migration

From over 1250 extant species of the order Chiroptera, 25 and 28 are known to occur in Germany and Hungary, respectively. Close to 350 samples originating from 28 bat species (17 from Germany, 27 from Hungary) were screened for the presence of adenoviruses (AdVs) using a nested PCR that targets the DNA polymerase gene of AdVs. An additional PCR was designed and applied to amplify a fragment from the gene encoding the IVa2 protein of mastadenoviruses. All German samples originated from organs of bats found moribund or dead. The Hungarian samples were excrements collected from colonies of known bat species, throat or rectal swab samples, taken from live individuals that had been captured for faunistic surveys and migration studies, as well as internal organs of dead specimens. Overall, 51 samples (14.73%) were found positive. We detected 28 seemingly novel and six previously described bat AdVs by sequencing the PCR products. The positivity rate was the highest among the guano samples of bat colonies. In phylogeny reconstructions, the AdVs detected in bats clustered roughly, but not perfectly, according to the hosts’ families (Vespertilionidae, Rhinolophidae, Hipposideridae, Phyllostomidae and Pteropodidae). In a few cases, identical sequences were derived from animals of closely related species. On the other hand, some bat species proved to harbour more than one type of AdV. The high prevalence of infection and the large number of chiropteran species worldwide make us hypothesise that hundreds of different yet unknown AdV types might circulate in bats.

Sign in / Sign up

Export Citation Format

Share Document