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Objectives: C Trichomoniasis is one of the most common sexually transmitted diseases in the world, which is caused by Trichomonas vaginalis. It is also the most commonly reported sexually transmitted disease after the viral infections, which affects around 180 million people around the world each year. The people infected with this parasite exhibit a wide range of symptoms. To the best of our knowledge the genetic variation, prevalence and related factors affecting the disease have not been well studied. Therefore, this study aimed to determine the prevalence of T. vaginalis in women of southeast of Iran. Materials and Methods: Out of 500 patient women referred to the hospitals of Imam Khomeini in Zabol and Ali Ibn Abi Talib (AS) in Zahedan, 25 positive clinical samples were isolated from vaginal discharge and urine by culture method during June 2015 and May 2016. First, DNA was extracted and then all samples were subjected to nested PCR. Six different genotypes of actin gene were identified by PCR-RFLP in Trichomonas vaginalis in Zahedan and Zabol. All PCR products were digested with HindII, RsaI, and MesI restriction enzymes. All participants completed a questionnaire recommended by gynecologists and midwifery experts. Results: As a result, the genotypes of H, G, E, I, and N were identified in this study, from which the genotype E was the dominant genotype of T. vaginalis in Zahedan and Zabol. There was also a significant association between the type of clinical symptoms and the level of infection (P=0.0001). Conclusions: To sum up, disease as a health problem must be controlled through epidemiologic and genetic methods. Moreover, controlling the disease is closely associated with education and drug resistance or sensitivity related to genetic variation and epidemiologic factors. [ABSTRACT FROM AUTHOR]

Actin is a multi-functional gene family that can be divided into muscle-type actins and non-muscle-type actins. In this study, 37 unigenes encoding actins were identified from RNA-Seq data of Pacific white shrimp, Litopenaeus vannamei. According to phylogenetic analysis, four and three cDNAs belong to cytoplasmic- and heart-type actins and were named LvActinCT and LvActinHT, respectively. 10 cDNAs belong to the slow-type skeletal muscle actins, and 18 belong to the fast-type skeletal muscle actins; they were designated LvActinSSK and LvActinFSK, respectively. Some muscle actin genes formed gene clusters in the genome. Multiple alternative transcription starts sites (ATSSs) were found for LvActinCT1. Based on the early developmental expression profile, almost all LvActins were highly expressed between the early limb bud and post-larval stages. Using LvActinSSK5 as probes, slow-type muscle was localized in pleopod muscle and superficial ventral muscle. We also found three actin genes that were down-regulated in the hemocytes of white spot syndrome virus (WSSV)- and Vibrio parahaemolyticus-infected L. vannamei. This study provides valuable information on the actin gene structure of shrimp, furthers our understanding of the shrimp muscle system and helps us develop strategies for disease control and sustainable shrimp farming. [ABSTRACT FROM AUTHOR]

In animals and land plants, many asexual species originate through inter- or intraspecific crosses, and such heterozygous asexuals frequently are more abundant than their sexual relatives in marginal habitats. Although asexual species have been reported in various macroalgal taxa, detailed information regarding their distribution, heterozygosity, and origin is limited. Because many asexual tetrasporophyte strains of Caloglossa vieillardii have been isolated from South Australia, far from their core tropical habitats, we re-examined the distribution range of asexual C. vieillardii and genotyped these and other western Pacific strains using an actin gene marker. We confirmed the marginal distribution of the asexuals; however, a small patch of sexual thalli was newly discovered 450 km further west from asexual populations in South Australia. Three heterozygous genotypes and one homozygous genotypes were detected from nine asexual populations; 21 heterozygous strains were obligately asexual, but one homozygous strain suddenly produced sexual gametophytes after several years of culture. We hypothesized that the most abundant heterozygous genotype (defined as type 3/4) in asexual populations occurred by a cross between type 3 and type 4 allele gametophytes, both of which were isolated from the Australian coasts. In the crossing experiments, certain combinations between type 3 females and type 4 males produced tetrasporophytes, which recycled successive tetrasporophytes. In the culture experiments, whereas both sexual and asexual strains successfully produced tetraspores at 12°C, no sexual strains released carpospores below 14°C. However, it is uncertain whether this slight difference of maturation temperature was related to the marginal distribution of asexual C. vieillardii. [ABSTRACT FROM AUTHOR]

Background: The aim of this study was to identify the Trichomonas vaginalis strains/haplotypes based on identifying their probable variations in asymptomatic patients referred to Tabriz health centers, northwestern Iran. Methods: Sampling was taken from 50-suspected women to T. vaginalis in northwestern Iran. The obtained samples were smeared and cultured. Fifty DNA samples were extracted, amplified and identified by nested polymerase chain reaction and PCR-RFLP of actin gene using two endonuclease enzymes: MseI and RsaI. To reconfirm, the amplicons of actin gene were directly sequenced in order to identify the strains/haplotypes. Results: PCR-RFLP patterns, sequencing and phylogenetic analyses revealed definitely the presence of the G (n=22; 73.4%) and E (n=8; 26.6%) strains. Multiple alignments findings of genotype G showed five haplotypes and two amino acid substitutions in codons 192 and 211 although, no remarkable unique haplotype was found in genotype E. Conclusion: The accurate identification of T. vaginalis strains based on discrimination of their unknown haplotypes particularly those which are impacted on protein translation should be considered in parasite status, drug resistance, mixed infection with HIV and monitoring of asymptomatic trichomoniasis in the region. [ABSTRACT FROM AUTHOR]

[Display omitted] • A specific real-time PCR protocol was developed for faster detection of H. costale. • In France, H. costale was detected in adult oysters since 2005 and spat since 2008. • The H. costale prevalence in spat (14.59%) appeared higher than in adults (6.50%). • Actin gene sequencing showed 2 H. costale strains based on 3 sequence mutations. • The strain 1 was found in hatchery spat while the strain 2 was found in wild spat. The parasite Haplosporidium costale is known to infect and cause mortality in the oyster Crassostrea virginica in the USA. Decades after its first description in the 1960s, this parasite was detected in Crassostrea gigas in the USA and China. However, it presented a low prevalence and no mortality was associated with it. More recently, in 2019, H. costale was detected in France in a batch of moribund oysters. In order to observe how long this parasite has been present on French coasts, from Normandy to Thau lagoon, a retrospective investigation was conducted on 871 adult and spat oyster batches from 2004 to 2020. To allow rapid detection on a large panel of samples, a real-time PCR for the H. costale actin gene was developed. This method allowed the detection of H. costale DNA in adults from 2005 and in spat from 2008. The H. costale prevalence in spat appeared higher than in adults over the years studied, 14.59 % compared to 6.50 %, respectively. All samples presenting positive results were then sequenced on two targets, H. costale rRNA and actin genes. The actin gene sequencing highlighted the presence of two H. costale strains. Adult C. gigas as well as spat batches coming from hatcheries and DNA controls from C. virginica all presented with the Profile 1 H. costale strain. The Profile 2 H. costale strain was detected only in C. gigas spat coming from natural sources. These observations suggest a correlation between the origin of oysters and H. costale strains which may have been caused by commercial imports between Japan, USA and France back to the 1970s. Over the positive samples studied, only few batches (n = 3) suffered mortalities which could be hypothesized to be caused by H. costale , all presenting the Profile 1 H. costale strain. [ABSTRACT FROM AUTHOR]

Cryptosporidium is an enteric parasite that infects humans and a wide range of animals. Relatively little is known about the epidemiology and taxonomy of Cryptosporidium in fish. In the present study, a total of 775 fish, belonging to 46 species and comprising ornamental fish, marine fish and freshwater fish were screened for the prevalence of Cryptosporidium by PCR. The overall prevalence of Cryptosporidium in fish was 5.3% (41/775), with prevalences ranging from 1.5 to 100% within individual host species. Phylogenetic analysis of these Cryptosporidium isolates as well as 14 isolates from previous studies indicated extensive genetic diversity as well as evidence for mixed infections. At the 18S locus the following species were identified; Cryptosporidium molnari- like genotype ( n = 14), Cryptosporidium huwi ( n = 8), piscine genotype 2 ( n = 4), piscine genotype 3-like ( n = 1), piscine genotype 4 ( n = 2), piscine genotype 5 ( n = 13), piscine genotype 5-like ( n = 1) and five novel genotypes ( n = 5). At the actin locus, species identification agreed with the 18S locus for only 52.3% of isolates sequenced, indicating high levels of mixed infections. Future studies will need to employ both morphological characterization and deep sequencing amplicon-based technologies to better understand the epidemiological and phylogenetic relationships of piscine-derived Cryptosporidium species and genotypes, particularly when mixed infections are detected. [ABSTRACT FROM AUTHOR]

Background: Trichomonas vaginalis is the agent of urogenital tract infection that causes human trichomoniasis with some serious health complications. More understanding about genetic features of the parasite can be helpful in the study of the pathogenesis, drug susceptibility and epidemiology of the infection. For this end, we conducted analysis of the actin gene of T. vaginalis by applying the PCR-SSCP (PCR-Single Stranded Conformational Polymorphism) and nucleotide sequencing method. Methods: Fifty T. vaginalis samples were collected from 950 women attending gynecology clinics in two cities of Iran, Hamadan and Tehran, from November 2010 to July 2011. After axenisation of isolates, all samples subjected to PCR-SSCP and nucleotide sequencing. Results: According to the SSCP banding patterns and nucleotide sequencing, seven sequence types were detected among the isolates. Alignment of the nucleotide sequences showed five polymorphic sites in the different strain types. Amino acid substitution was not observed in the nucleotide sequence translation of the all sequences. Conclusion: The actin gene analysis represents genetic diversity of T. vaginalis and it suggests that various strains can be responsible for clinically different trichomoniasis in infected individuals. It is expected that further studies will be conducted to increase our knowledge about relationship between the actin gene polymorphism and different biological behavior of the parasite. [ABSTRACT FROM AUTHOR]

This study describes the isolation, cloning and phylogenetic studies of a partial sequence of actin gene from the fungus Trichoderma harzianum E-58, which was grown in Vogel's medium at 28°C and pH 5.5, with glucose as a carbon source. For the amplification of b-actin gene, the genomic DNA was subjected to the polymerase chain reaction (PCR) by using sequence specific primers. Through agarose gel electrophoresis the amplified PCR product was purified and ligated into pTZ57R/T vector. The ligation mixture was then transformed into E. coli DH10B and spread on ampicillin containing LB agar plate. Restriction analysis was done to confirm the positive transformants. Further, the gene was sequenced, and on the basis of this partial sequence, a dendrogram was made to assess homology among different fungi. These results will surely help study the regulation of cellulase gene expression in the fungus in future. [ABSTRACT FROM AUTHOR]

Human trichomoniasis, caused by the protozoan Trichomonas vaginalis, is a highly prevalent sexually transmitted infection. However, little is known about the degree of strain variability of T. vaginalis. A reliable classification method for T. vaginalis strains would be a useful tool in the study of the epidemiology, pathogenesis and transmission of T. vaginalis. A PCR–restriction fragment length polymorphism typing method was designed and evaluated using T. vaginalis isolates obtained after culture of vaginal specimens collected in the Democratic Republic of Congo and in Zambia. The variation of the actin gene of T. vaginalis was determined for three ATCC reference strains and 151 T. vaginalis isolates. Eight different types were identified, on the basis of the digestion patterns of the amplified actin gene, with each of the restriction enzymes HindII, MseI and RsaI . It was determined that the ATCC reference strains 30001, 30240 and 50141 were of actin genotypes G, H and E, respectively. The actin genotype type E was more common in the Democratic Republic of Congo, whereas type G was the commonest type in Zambia. Translation of the nucleotide sequence showed up to three amino acid substitutions. We developed a reproducible, sensitive and specific typing method for T. vaginalis, and were able to distinguish at least eight T. vaginalis actin genotypes. Further studies are needed to evaluate the method using clinical specimens and to determine the utility of the typing method for the genotypic characterization of T. vaginalis. [ABSTRACT FROM AUTHOR]

Bonamia ostreae is a protozoan parasite that infects the European flat oyster Ostrea edulis, causing systemic infections and resulting in massive mortalities in populations of this valuable bivalve species. In this work, we have characterized B. ostreae actin genes and used their sequences for a phylogenetic analysis. Design of different primer sets was necessary to amplify the central coding region of actin genes of B. ostreae. Characterization of the sequences and their amplification in different samples demonstrated the presence of 2 intragenomic actin genes in B. ostreae, without any intron. The phylogenetic analysis placed B. ostreae in a clade with Minchinia tapetis, Minchinia teredinis and Haplosporidium costale as its closest relatives, and demonstrated that the paralogous actin genes found in Bonamia resulted from a duplication of the original actin gene after the Bonamia origin. [ABSTRACT FROM PUBLISHER]

Abstract: An actin gene (named Mar-actin) from the commercial prawn, Macrobrachium rosenbergii, was isolated, sequenced and gene expression was characterized. The cDNA sequence was 1281 bp in length and contained 1131 bp open reading frame encoding 376 amino acids. The amino acid sequence deduced from the nucleotide sequence showed high identity (70.3% to 98.1%) with other known actins of various organisms, highest with that of the European flounder (98.1%). The 3′ untranslated region (3′ UTR) of the Mar-actin mRNA has a high A+U content (approximately 78%) and contains one AUUUUUA and two repeats of the AUUUA motifs, that may function in regulating mRNA decay. Northern blot analysis revealed that the Mar-actin gene was expressed predominantly in muscle tissues. Transcripts in hepatopancreas were barely detectable. Expression of the Mar-actin gene varied during embryonic development and reached the maximal level at the zoea stage. This is the first report describing the complete sequence and expression pattern of the actin gene in prawns. [Copyright &y& Elsevier]

Highly similar gene sequences of the 5′ region of the large subunit (LSU) are commonly interpreted to predict the organism''s identity. However, it was recognised that closely related taxa do not always show sufficiently diverged D1/D2 LSU sequences to differentiate between them. The effectiveness of species separation using D1/D2 LSU sequences, small subunit (SSU) sequences and actin gene sequences was determined by pair-wise comparisons. The LSU data showed coinciding similarities among and within species. The actin data resolved all investigated species. Examples strengthened the value of almost complete SSU sequences for species separation. The larger number of differences in the highly conserved actin gene, compared to the overall more variable LSU gene, is due to the tolerance of protein coding genes to synonymous nucleotide changes. In contrast, the pairing in secondary structures of the rRNA, ensuring the functionality of the molecule, relies on longer and uninterrupted sequence sections. In conclusion, D1/D2 LSU sequences are not specific enough to identify closely related taxa. The actin gene is a better marker in these cases. However, because of the availability of a large database of fungal D1/D2 LSU sequences, this gene region is currently still the preferred target for sequence-based identification. [Copyright &y& Elsevier]

A novel one-step method for the differentiation of chicken and turkey DNA is described. The technique uses the polymerase chain reaction (PCR) and primers that exploit intron variability in α-cardiac actin to generate single products of a characteristic size for each species. No cross-reactivity with porcine, ovine or bovine DNA templates is apparent and analysis of chicken/turkey admixtures indicates that it is possible to detect 1% turkey in 99% chicken and vice versa. Because the test is based on a nuclear gene target, it forms a valuable complement to other methods based on mitochondrial DNA sequences. [Copyright &y& Elsevier]

Regulation of the vertebrate actin multigene family involves the recognition of various regulatory sequences (cis-acting elements) that specify the distinct tissue type and developmental program of expression for each actin paralogue, which implies that the distribution of cis-acting elements may be unique for each paralogue gene. To elucidate the evolution of these unique distribution patterns, we improved a method to scan for cis-acting elements in the 5′ flanking regulatory region of genes and used it to analyze five cis-acting elements (SRE, MyoD binding site, Elk-1 binding site, positive and negative YY1 binding sites) of six actin paralogue genes (β and γ cytoplasmic actins, α and γ smooth muscle actins, and α skeletal and α cardiac actins) among various vertebrates. It was shown that although an element(s) may exist in all paralogue genes of the same species, its numbers, compositions, and distribution patterns or even sequences vary remarkably among paralogues, which contributes to their different tissue- and developmental-specific expression. However, each pair of coexpressed paralogues has some certain similarity in distribution patterns. Furthermore, among various orthologues of actin genes derived from diverse vertebrates, the sequences, numbers, and distribution patterns of these cis-acting elements are highly conserved or even identical in the long run of phylogeny of vertebrates. Taken together, the results described above strongly indicate that not only the structures of actins but also their expression patterns are essential in both the phylogeny and the physiology of vertebrates. The distribution patterns of cis-acting elements of various actin genes can be regarded as indicators of both horizontal (paralogous) and vertical (orthologous) evolution of actins. [ABSTRACT FROM AUTHOR]

This paper reports the cloning and sequencing of a Phytophthora megasperma actin gene. Sequence analysis revealed that the gene does not contain introns and encodes a putative actin protein remarkably diverged from any other known actins. Genomic Southern analysis indicates that there is a single actin gene per haploid genome, which is actively transcribed in vivo, as revealed by S1 mapping of the transcripts. The transcripts are heterogeneous in size because they have different 3′ termini, none of which contains the conventional AATAAA polyadenylation signal. [ABSTRACT FROM AUTHOR]