Paweł Łyp, Michał Bartnicki, Marta Staniec, Stanisław Winiarczyk and Łukasz Adaszek
.: Canine babesiosis in Romania due to Babesia canis and Babesia vogeli: a molecular approach. Parasitol Res 2012, 110, 1659-1654.
12. Kostro K., Stojecki K., Grzybek M., Tomczuk K.: Characteristics, immunological events, and diagnostics of Babesia spp. infection, with emphasis on Babesia canis. Bull Vet Inst Pulawy 2015, 59, 495-504.
13. Łyp P., Adaszek Ł., Furmaga B., Winiarczyk S.: Identification of new 18SrRNA strains of Babesia canis isolated from dogs with subclinical babesiosis. Pol J Vet Sci 2015, 18, 573-577.
Entamoeba gingivalis normally exists in the human oral cavity, namely in the gums, and brings about some specific diseases. However, it can also trigger some more serious illnesses. Among these are infections of the genital tract, acute osteomyelitis of the mandible and pulmonary abscess. Entamoeba gingivalis identification by light microscopy is difficult, hence polymerase chain reaction (PCR) is used. The contemporary primers for PCR are complement to 18S rRNA. This article informs the reader of the process that was involved in designing new primers for three genes which were thought to be present on the Entamoeba gingivalis genome, but their sequences were unknown. The newly obtained sequences of primers have better properties for identification purposes, compared to these which are currently used.
K. Quiazon, T. Yoshinaga, H. Doi, J. Araki and K. Ogawa
Finding male philometrid nematodes is essential for taxonomic identification among congeneric species. In this study, male Philometra thaiensis Moravec, Fiala et Dyková, 2004 were collected and described for the first time, from the body cavity of the freshwater fish (eyespot pufferfish) Tetraodon biocellatus Tirant (Tetraodontiformes, Tetraodontidae), and conspecific females were redescribed based on the additional morphological biometrics examined. Molecular examination was carried out on the small subunit 18S rRNA, revealing the evolutionary relationships of P. thaiensis and reported philometrid species (Philometra and Philometroides) from Japan with other dracunculoids deposited in the GenBank. Based on the molecular data, there are some genera (Philometra, Philometroides, Clavinema, and Margolisianum [genus inquirendum]) requiring further morphological re-evaluation that should be supported with molecular data.
Two species of Thaparocleidus Jain (1952a) were found harboring W. attu from the Ganga River at two localities, Meerut and Farrukhabad, Uttar Pradesh, India, during the period of 2013-2015. Morphology and morphometric study of specimens identified as Thaparocleidus gomtius (Jain, 1952a) Lim, 1996 and T. sudhakari (Gusev, 1976) Lim, 1996. Molecular analyses using the 18S rRNA gene confirmed the validity of T. gomtius and T. sudhakari and demonstrated that both the species clustered with other Thaparocleidus species from different geographical regions. We aim at reassessing the taxonomy and establishing the phylogenetic relationships among these two redescribed species with other representatives of the genus Thaparocleidus.
A new host and new locality is recorded for Diplostamenides sciaenae (Goto, 1894) Lebedev, Parukhin et Roitman, 1970 from Johnius belangerii at Versova dock landing centre, Mumbai, India. The morphometric comparison of D. sciaenae with previously published data, provided redescription complements results of molecular analysis. The partial 28S and 18S rRNA gene sequences of D. sciaenae were amplified, sequenced through PCR and deposited to GenBank database. The BLASTn searches revealed the significant closeness of D. sciaenae to other microcotylid parasites in large and small ribosomal subunits. Th e phylogenetic tree analyses with neighbor joining and minimum evolution methods also expressed belonging of D. sciaenae to Microcotylidae.
Cryptosporidium infections has been reported in several avian species including chickens, pigeons and game birds where these infections had been identified to cause either enteric or respiratory diseases. However, little data exists on the molecular characterization of Cryptosporidium species in ducks, especially those in frequent contact with humans. The aim of this study was to detect the Cryptosporidium species infecting domestic ducks in two major live bird markets. A total of 109 fresh faecal samples were collected from all the ducks available on sale in the two markets. The detection of Cryptosporidium species was conducted by microscopy. All positive samples were confirmed by the nested PCR amplification and the nucleotide sequencing of the 18S rRNA genes. The results demonstrated that the prevalence of Cryptosporidium infection in ducks using microscopy was 11.0 % (12/109). There was a higher prevalence 14.0 % (7/50) in ducks from Ibadan compared with those 8.5 % (5/59) obtained from Oyo town. All positive samples by microscopy were also positive using the nested PCR and the DNA sequencing of the secondary PCR products from the 18S rRNA genes which revealed the presence of Cryptosporidium parvum. This study revealed that natural infections of C. parvum may occur in ducks in close contact with humans and other domestic animals and therefore suggests that cryptosporidiosis in ducks may be of public health importance.
Reference genes are generally used as endogenous normalization factor for relative quantification of target genes in quantitative real-time PCR (qRT-PCR). The present work aimed at identifying suitable reference genes for normalization of qRT-PCR data in tissues of Eucalyptus tereticornis. The expression levels of housekeeping genes like Actin (EtAct2), Isocitrate dehy - drogenase (EtIDH), ribosomal RNA (Et18s rRNA), SAND family protein (EtSAND), Histone protein (EtH2B), α-Tubulin (EtTUB), and eukaryotic initiation factor (EteIF4B) were studied to characterize their normalization stability in different tissues including young leaves, internodes, developing and mature xylem. The expression level of these genes was analyzed using different algorithms like geNorm, NormFinder and Best- Keeper. Among the seven reference genes analyzed, EtAct2 was expressed with less variance and was found to be the most stable reference gene across different tissues using all the three programs, while the least stable gene identified was EtH2B. Further, the normalization efficiency of the reference genes were assessed to predict the expression levels of three primary cell wall specific cellulose synthase transcripts (EtCesAs) in E. tereticornis tissues. The relative expression of EtCesA4, EtCesA5 and EtCesA6 was determined to be 3-19 fold higher in leaf and internode tissues when compared to developing and mature xylem tissues. This study will allow accurate normalization of qRT-PCR experiments across different tissues in E. tereticornis for future genomic research in this tropical eucalypt species.
Joanna Pławińska-Czarnak, Joanna Zarzyńska, Janusz Bogdan, Alicja Majewska, Marek Karwański, Magdalena Kizerwetter-Świda, Jarosław Kaba, Krzysztof Anusz and Emilia Bagnicka
The goat (Capra hircus) is a perfect animal model for analyzing the transcriptome of milk somatic cells (MSCs), as sufficient numbers of somatic cells in goat milk, i.e., exfoliated epithelial cells, can be obtained using noninvasive methods. RNA integrity and purity are the first and most important parameters qualifying samples for transcriptomic tests and next-generation sequencing, as RNA quality influences experimental results. The aim of this study was to optimize a method for obtaining high-quality RNA from goat MSCs, irrespective of effects like breed, lactation stage, health status (e.g., with or without small ruminant lentivirus [SRLV] infection), or number of somatic cells. Milk samples were obtained from goats of two Polish breeds in various lactation stages and in different parities, and from goats infected and not infected with SRLV. Altogether, 412 MSC samples were examined: 206 using method A with fenozol and 206 using method B with QIAzol. Though the overall purity (measured as absorbance ratios at 260 nm/280 nm and 260 nm/230 nm) of the RNA material was comparable, the average yield of RNA isolated using method A was 11.9 µg, while method B’s average yield was 29.9 µg. Moreover, method B resulted in good quality RNA suitable for transcriptome analysis. Results were confirmed by RT-qPCR, using 18S rRNA and RPLP0 as the reference genes. The application of our modified treatment method was successful in obtaining high-integrity samples for transcriptomic or next-generation sequencing analysis. Using a 400 mL milk sample cooled in ice directly after milking, securing the cooling chain process from milking to MSC isolation, and applying method B to isolate RNA, we obtained good RNA quality irrespective of the goats’ breed, lactation stage, parity, milk yield, SRLV infection, and even milk yield and number of somatic cells in milk.
V. V. Besprozvannykh, K. V. Rozhkovan, A. V. Ermolenko and A. V. Izrailskaya
Trematoda . Volume 2. London, UK: CABI Publishing and The Natural History Museum, pp. 319 – 324
K rieger , J., H ett , A.K., F uerst , P.A., B irstein , V.J., L udwig , A. (2006): Unusual intraindividual variation of the nuclear 18SrRNA gene is widespread within the Acipenseridae. J. Hered. , 97, 218 – 225. DOI:10.1093/jhered/esj035
K uzmin , S.L. (2012): [ Amphibians of former USSR ] Moscow, Fellowship of scientific books. 370 p. (In Russian)
L ang , A. (1892): Über die Cercariae von Amphistomum subclavatum , Ber. Naturforsch. Ges. Freiburg im