Partial DNA Sequences Research Articles

Double trouble. A cryptic first record of Berghia marinae Carmona, Pola, Gosliner, & Cervera 2014 in the Mediterranean Sea

AbstractIn 2014, Berghia marinae Carmona, Pola, Gosliner & Cervera, 2014 from Senegal was described along with the revision of the genus Berghia Trinchese, 1877. In this study, we establish a second record for the senegalese species B. marinae in the Mediterranean Sea, 4,000 Km away from its type location. The morphological mismatch from the original description hampered its identification, and thus, a molecular approach was needed. Multilocus phylogenetic trees were inferred from Maximum-likelihood and Bayesian analyses based on partial DNA sequences of the mitochondrial cytochrome c oxidase subunit I and 16S rRNA genes, and the nuclear gene histone-3. Species delimitation analyses were performed to support the phylogenetic results and a new morphological description is provided complementing earlier information on this barely known species.

Native and introduced Far Eastern daces, genus Tribolodon, in the Tama River system, Japan, inferred from mitochondrial DNA analysis

To investigate the species composition and genetic characteristics of Far Eastern daces, genus Tribolodon, in the Tama River, Japan, partial DNA sequences of the mitochondrial DNA control region were compared among specimens from the whole river system. Specimens of Tribolodon hakonensis (n = 257) and Tribolodon brandtii maruta (n = 193) were examined in this study. Inconsistencies in species identification according to genetic and species-specific morphological traits, i.e., breeding color and cephalic lateral line system, were found between some sympatrically distributed specimens (17/450; 3.8%), indicating possible hybridization between T. hakonensis and T. brandtii maruta. The two species were sympatrically distributed in the lower reaches, whereas T. hakonensis solely appeared in the upper reaches and headwaters. In T. hakonensis, we found obvious genetic divergence between the lower and upper reaches, suggesting reproductive isolation related to alternative migration patterns of anadromous and river-resident forms, respectively. Further, it is suggested that native and introduced T. hakonensis populations coexist in the lower reaches and headwaters, whereas the former solely inhabit, or are dominant, in the upper reaches.

Cryptic Diversity in the Aphid-Parasitizing Wasp Protaphidius nawaii (Hymenoptera: Braconidae): Discovery of Two Attendant-Ant-Specific mtDNA Lineages.

The parasitoid wasp Protaphidius nawaii parasitizes the aphid Stomaphis japonica, which is obligatorily attended by several species of ants of genus Lasius. Subgenus Lasius or Dendrolasius ants use different defense strategies to protect the aphids that they attend (Lasius, shelter building; Dendrolasius, aggressive attack). We performed molecular phylogenetic analysis based on partial mitochondrial DNA sequences of P. nawaii and found that the parasitoid wasp consists of two highly differentiated genetic lineages. Although these two lineages distributed sympatrically, one tends to parasitize aphids attended by ants of subgenus Lasius, and the other parasitizes aphids attended by ants of subgenus Dendrolasius. The two lineages of P. nawaii appear to exhibit different oviposition behaviors adapted to the different aphid-protection strategies of the two ant subgenera.

Novel myxosporean species parasitizing an economically important fish from the Amazon basin.

This paper provides morphological and phylogenetic analyses of two new myxobolid species found infecting Piaractus brachypomus from the Amazon basin. The fish were caught in the Tapajós River, in the municipality of Santarém, in the state of Pará, Brazil. The plasmodial development of Henneguya brachypomus n. sp. occurred in the gill lamellae while Myxobolus pirapitingae n. sp. developed in the pyloric cecum. Morphological analyses did not identify inflammatory infiltrate for either species, but H. brachypomus n. sp. induced stretching of the epithelium, compression of the adjacent tissues, and displacement and deformation of the neighboring lamellae. The mature myxospores of H. brachypomus n. sp. were ellipsoid, with a length of 11.7-13.8μm, a width of 4.0-4.6μm, and a thickness of 3.5-4.3μm. The polar capsules were elongated, with a length of 5.6-7.3μm and a width of 1.3-2.0μm, and each contained a polar filament with 8-9 coils. The caudal process was 40.5-48.1μm long and the total length of the myxospore was 52.4-61.6μm. Myxobolus pirapitingae n. sp. exhibited rounded mature myxospores measuring 10.0-11.1μm in length, 7.0-7.6μm in width, and 5.4-6.3μm in thickness. The polar capsules were of equal size and occupied less than half the myxospore, measuring 3.5-4.0μm in length and 2.0-2.6μm in width, with each containing a polar filament with 6-7 coils. Phylogenetic analysis based on partial small subunit ribosomal DNA (ssrDNA) sequences showed that H. brachypomus n. sp. clustered as a sister species of Henneguya piaractus, while M. pirapitingae n. sp. was grouped in a sub-clade together with Myxobolus matosi and Myxobolus colossomatis.

Morphological Description, Phylogenetic and Molecular Analysis of Dirofilaria immitis Isolated from Dogs in the Northwest of Iran

Dirofilariasis is a globally distributed arthropod-borne parasitic disease of mainly canids and felids. We evaluated to extend the knowledge of morpho-molecular characteristics and outer ultrastructure of Dirofilaria immitis isolated from Northwest of Iran. Overall, 67 filarial worms including 41 females and 26 males parasites were collected from the cardiovascular system of the 43 stray dogs in Meshkinshar, Ardebil Province, Northwest of Iran in 2017, and subjected to light and scanning electron microscopy (SEM) as well as carmine alum staining for morpho-molecular and identification. Molecular methods were used for confirmation of morphological findings by sequencing of Cyto-chrome c oxidase subunit I (cox1) gene. The partial DNA sequencing of cox1 gene of adult parasites showed considerable homology and close proximity to the previously isolated from Kerman and Meshkinshahr, Iran. The lowest genetic variation and the highest intra-species variability was found in D. immitis and Dirofilaria repens, respectively. No similarity was identified between D. immitis nucleotide sequence and Wolbachia species as its endosymbiont bacteria. The SEM technique is an excellent tool for differential recognition of the parasite surface morphology and molecular techniques could differentiate and identify Dirofilaria spp.

Novel Single Nucleotide Polymorphisms of Lysozyme (C-Type) Gene in Donkey (Equus Asinus) Populations in Marmara Province of Turkey

The major antimicrobial proteins in donkey milk are lysozyme, lactoferrin, lactoperoxidase and immunoglobulins. Lysozyme has an important role in the host defense by way it inhibits the pathogenic bacteria. The aim of this study is to investigate the Lysozyme (LYZ) gene polymorphism in 82 donkeys reared in Thrace region of Turkey. 716 bp long partial 5‘ UTR, exon 1, intron 1, exon 2 regions of LYZ gene were amplified and PCR products were analyzed via DNA sequencing. Three novel single nucleotide polymorphisms (SNPs) were identified as g.1782775A>G, g.1782924A>G and g.1782960T>C in the first intron of LYZ gene. The partial DNA sequence of LYZ gene in donkeys was reported in the present study and sequences of LYZ were entered to NCBI Genbank database with the accession number: MK984689-MK984692. This SNP may have an effect on immune system and milk traits in donkeys and additional studies are needed to confirm this assumption for donkey breeding.

Genetic analysis and molecular phylogeny of zigzag leafhopper Maiestas dorsalis (Motschulsky) using mitochondrial COI gene

The zigzag leafhopper Maiestas dorsalis (Hemiptera) is a tungro virus vector that cause damage on rice plants. Genetic analysis of M. dorsalis that be isolated from Samosir island, North Sumatra, Indonesia using partial DNA sequence of mitochondrial cytochrome oxidase subunit I (COI) DNA is still limited. This study aims to identify and to find out the genetic data of M. dorsalis that be derived from region of its mitochondrial cytochrome oxidase I gene (COI). Analysis was done by four steps, namely: Hopper collection on paddy field by using insect net, DNA extraction by using Zymo Tissue & Insect DNA Mini Preparation, amplification by PCR using My TaqTM HS Red Mix and DNA sequence analysis using ABI PRISM 3730xl Genetic Analyzer. Primer cocktail tRWF-Mlep was used in DNA amplification step. The research result pointed out that COI DNA fragment of M. dorsalis has length 521 bp. This COI DNA sequence was dominated by A and T(U) bases with concentration 74.30%. The concentration of T(U), C, A and G nucleotides in the COI sequence were 35.90%, 13.40%, 38.44%, and 12.30%, respectively. Identification of M. dorsalis based on this COI DNA sequence confirmed the identification result based on its morphological characters.

First record of Ahaetullamycterizans (Linnaeus, 1758) (Serpentes: Colubridae) from the Lesser Sunda region, Indonesia, based on molecular and morphological identification

Previous studies on the distribution of the Ahaetulla snake across Indonesia only focused on morphological characters without any molecular data. This study was aimed at analyzing phylogenetic relationships among the genus Ahaetulla in Indonesia based on partial mitochondrial DNA sequences of 16 specimens collected from the Sundaland and Lesser Sunda regions. The 12S-rDNA gene was PCR-amplified and sequenced to analyze phylogenetic relationships and to estimate the divergence time between the 2 Ahaetulla populations of the Sundaland and Lesser Sunda regions. Morphological characters of 3 Ahaetulla specimens from Lesser Sunda and Sundaland have also been analyzed to confirm the results based on molecular markers. Both the phylogenetic analyses and morphological characters revealed the presence of 2 Ahaetulla species, A. mycterizans and A. prasina, in Sundaland as well as in the Lesser Sunda region. Previously, A. mycterizans was only known to exist in Sundaland. Its presence in the Lesser Sunda region is reported here for the first time. The divergence time estimations suggested the presence of A. mycterizans in Lesser Sunda had occurred during the Miocene, around 12.1 Ma. Consequently, this study has added A. mycterizans to the list of snakes that inhabit Lesser Sunda and revealed the need to initiate a survey in more detail to investigate the presence of A. mycterizans in other regions of Southeast Asia.

Report on a new species of Plutomurus (Collembola, Tomoceridae)from a South Korean limestone cave, with notes on its DNA barcode.

A new species of Plutomurus Yosii, 1956, P. jangamensis sp. nov., was found in a limestone cave of South Korea. The new species is very similar to P. gul (Yosii, 1966) in the general pattern of cephalic and body chaetotaxy, absence of eye, presence of pointed tenent hair, number of lateral macrochaetae in basal segment of dens, and number of mucronal intermediate tooth; however, the two species can be distinguished by the small spines arranged in-a-row along with dental spines and lighter body color. This species, however, shows variability and was not easily classified morphologically as P. jangamensis sp. nov. and P. gul., in terms of body color, formula of dental spine, or even the number of prelabral chaetae. Partial DNA sequences of the mitochondrial cytochrome c oxidase subunit I (COI) gene were used as DNA barcodes to distinguish the species. This result suggests that the COI gene is useful for discrimination of P. jangamensis sp. nov. with high support (bootstrap=100). An identification key to the world species of Plutomurus is also provided.

Identification and Characterization of Colletotrichum Species Associated with Anthracnose Disease of Camellia oleifera in China.

Tea-oil tree (Camellia oleifera Abel) is an important edible oil woody plant with a planting area over 3,800,000 hectares in southern China. Anthracnose is a serious disease of tea-oil tree in southern China, causing severe economic losses and posing a huge threat to the Ca. oleifera industry. Based on recent developments in the classification of Colletotrichum species, the objective of this study was to identify Colletotrichum species associated with tea-oil tree and examine their pathogenicity on leaves and fruits of Ca. oleifera. In total, 232 isolates were obtained from Ca. oleifera leaves and fruits with anthracnose symptoms. These isolates were further characterized based on morphology and multilocus phylogenetic analyses using partial DNA sequences at the ribosomal internal transcribed spacer regions and β-tubulin, actin, calmodulin, glyceraldehyde-3-phosphate dehydrogenase, and chitin synthase-encoding genes. The fungal isolates belong to five species: C. camelliae, C. fructicola, C. siamense, C. aenigma, and C. gloeosporioides. C. camelliae was the most predominant and widely distributed species on fruits of Ca. oleifera (91.4%), followed by C. fructicola (6.3%). However, C. fructicola was common and widely distributed species on leaves (75.9%), followed by C. camelliae (17.2%). There was no evidence of geographical specialization of the different species. Pathogenicity assays showed that all tested isolates, including 20 of C. camelliae, 11 of C. fructicola, four of C. siamense, two of C. aenigma, and one of C. gloeosporioides, were pathogenic to leaves and fruits of Ca. oleifera. Among the five species, C. camelliae species showed strong pathogenicity on both leaves and fruits of Ca. oleifera, and C. fructicola, C. siamense, C. aenigma, and C. gloeosporioides all showed weak pathogenicity on both leaves and fruits. No relationship was found between origin of isolates and their virulence. This is the first description of C. camelliae, C. fructicola, C. siamense, and C. gloeosporioides from the fruits of Ca. oleifera in China.

Association of HSF1 Genetic Variation with Heat Tolerance in Chinese Cattle.

Simple SummaryChina harbors two lineages of cattle (Bos taurus and Bos indicus). North China bears Bos taurus. Bos indicus predominantly belong to south China, whereas in central China there is an admixture of Bos taurus and Bos indicus, representing a discontinuity with north and south China. It has been reported that Bos indicus are more heat resistant than Bos taurus and heat stress can damage the structure of protein. Heat shock factor1 (HSF1) is a candidate heat tolerance gene which works by increasing expression of nascent heat shock proteins (HSPs), promoting the refolding of denatured proteins. Conversely, despite its importance, the effects of the bovine HSF1 gene on heat tolerance traits in Chinese cattle remain unknown. Our results suggest that a missense mutation of the HSF1 gene is associated with the heat tolerance capability of Chinese cattle.The heat shock factor 1 (HSF1) gene is a regulator of the heat stress response, maximizing HSP protein expression survival. In this research, we explored the frequency distribution of a missense mutation (NC_037341.1 g.616087A > G, rs135258919) in the HSF1 gene in Chinese cattle with amino acid substitution, valine to alanine. This mutation could be related to the heat tolerance in Bos indicus. A total of 941 individuals representing 35 Chinese native cattle breeds, combining pure taurine (Angus) and indicine cattle, were used to determine the genotypes of the mutation through PCR and partial DNA sequencing. The results showed significant differences in allele frequencies and their genotypes amongst Chinese cattle from different regions. Allele G or indicine-specific allele frequency diminished from south to north China, while allele A (genotype AA) or the taurine-specific allele had a contrary pattern, which agreed with the distribution of taurine and indicine cattle. According to the association analysis, the NC_037341.1 g.616087A > G (rs135258919) of the bovine HSF1 gene, annual temperature (T), relative humidity (RH), and the temperature humidity index (THI) (p < 0.01) were interrelated closely, which indicated that the NC_037341.1 g.616087A > G of the HSF1 gene is associated with heat tolerance in indicine cattle.

Effect of time of therapy with wild-type lytic bacteriophages on the reduction of Salmonella Enteritidis in broiler chickens

Salmonella Enteritidis remains a leading cause of human foodborne disease, mostly associated with the consumption of contaminated poultry products. To more strategically implement a phage therapy scheme for S. Enteritidis control in broilers, a cocktail containing three wild-type lytic bacteriophages (LBs) previously isolated from chickens was evaluated shortly and later after a challenge. Genomic characterization, lytic spectrum and in vitro efficacy were determined for each studied LB. In independent trials, broilers challenged with S. Enteritidis on day of hatch received phage therapy from 6 to 10 days of age (early treatment), and from 31 to 35 days of age (later treatment). S. Enteritidis analyses were performed before treatment and at 1, 4, 7 and 10 days post-treatment (dpt) in both trials. Partial DNA sequence analysis of each LB revealed close similarity to the Ackermannviridae family. LBs lysed different Salmonella enterica serovars, while other tested bacteria were refractory. An in-vitro reduction of 1.49, 0.65 and 0.58 log10 CFU/mL in S. Enteritidis number was obtained after co-incubation for 3 h with each LB. Both in vivo trials showed a significant reduction in the average number of intestinal S. Enteritidis calculated after phage therapy compared with controls. However, the highest efficiency was found in the later therapy, which resulted in a reduction of 1.08 log10 CFU/g in the average from 4 to 10 dpt, showing potential for future use as a pre-harvest strategy to reduce the S. Enteritidis intestinal colonization in broilers on farms.

Leptin Receptor Gene Polymorphisms in Some Turkish Donkey Populations

Leptin receptor is a fundamental regulator in physiological functions of the regulation of food intake, energy homeostasis, immune function, and reproduction as well as on ovarian follicular cells on the placenta and lactating mammary glands. The aim of this study was to investigate the LEPR gene polymorphism in 60 donkeys reared in Thrace region of Turkey. A 585 bp long partial intron 6, exon 7, intron 7, and exon 8 regions of LEPR gene were amplified, and polymerase chain reaction products analyzed via DNA sequencing. A novel single-nucleotide polymorphism (SNP) was identified as g.713668A>G in the seventh exon region of LEPR gene. This novel SNP was first identified, and the partial DNA sequence of LEPR gene in donkeys was reported for the first time in this study, and these sequences were deposited to NCBI Genbank database with the accession number: MK807114-MK807115. The A>G transition revealed a silent mutation (CAA-CAG) in glutamine amino acid. This nucleotide mutation could cause the changes of secondary structure of protein and expression level of LEPR hormone. For this reason, additional studies are needed to reveal new SNPs and in the LEPR gene that may affect economic traits and structure of protein in donkey breeds.

First Report Regarding the Simultaneous Molecular Detection of Anaplasma marginale and Theileria annulata in Equine Blood Samples Collected from Southern Punjab in Pakistan.

The present study was designed to check the molecular detection of Anaplasma marginale and Theileria annulata in blood samples of horses and donkeys collected from Dera Ghazi Khan District in Punjab and to document their phylogenetic origin and their association with studied epidemiological factors (sex and age) and complete blood count parameters, if any. A total of 195 blood samples were collected from apparently healthy horses (N = 141) and donkeys (N = 54). A. marginale DNA was detected by PCR in 4.9% (7/141) horse and in 9.2% (5/54) of donkey blood samples. Prevalence of T. annulata was 5.6% (8/141) and 11.1% (6/54) in horse and donkey samples, respectively. While 1.4% (N = 2) horses and 3.7% (N = 2) donkeys were found co-infected with both parasites. Representative amplicon for both parasites was confirmed by DNA sequenced and partial DNA sequence of the major surface protein-1b encoding gene of A. marginale and cytochrome b gene from T. annulata were submitted to the GenBank database under the accession number MK792344-MK792348. Epidemiological data analysis revealed that female horses were more prone to A. marginale (P = 0.02) while female donkeys were more susceptible to A. marginale (P < 0.001) and T. annulata (P < 0.001) infection. It was observed that horse and donkey infected either with Anaplasma marginale or Theileria annulata had significantly disturbed red and white blood cell counts and their associated parameters. This is a first ever study regarding molecular detection of A. marginale and T. annulata in equine blood samples from Pakistan. We recommend that this multiplex PCR protocol should be used for the detection of Anaplasma marginale and Theileria annulata in livestock for their proper diagnosis and treatment.

The DNA Sequence Encoding Actin (ACT1) of Pandan (Benstonea sp.)

One of housekeeping genes is actin gene. This gene is frequently used in gene expression studies as an internal control. The DNA sequence encoding actin from Pandan ( Benstonea sp.) clone Riau has not been reported, therefore, this study investigated the DNA sequence encoding actin isolated from Benstonea sp. clone Riau. Total DNA isolation was performed in fresh leaves, total RNA isolation from stem, total cDNA synthesis, polymerase chain reaction using degenerate actin primer, electrophoresis, cloning, transformation, blue white colony selection, colony PCR, sequencing, data analysis using BioEdit and MEGA6 softwares and BLASTn program. The partial DNA sequence encoding actin from Benstonea sp. clone Riau obtained was 1,403 bp. The sequence was grouped as part of actin1 (ACT1) and it was consisted of two exons and one intron. The predicted coding and peptide sequences were 616 bp and 205 amino acids, respectively. The predicted coding sequence had 90% similarity to some ACT1 mRNA from some plants species but none of which belongs to Benstonea genus or Pandanaceae family. The deduced peptide sequence had similarity to some ACT1 peptide from some plant species of up to 99% and also none of them belongs to Benstonea genus or Pandanaceae family. Thus, the partial ACT1 gene obtained in this study was the first sequence reported from Benstonea genus. Furthermore, this sequence can be used as a reference to isolate actin genes from other species within Benstonea genus for gene expression analysis purposes.

1348. Mycobacterium Chimera Infection Following Cardiac Surgery: A Review of a Large Cohort of Cases in the United States

BackgroundMycobacterium chimera (MC) is a nontuberculous mycobacterium associated with infections originating from heater-cooler devices following cardiac surgery globally from 2012 to 2016. Twenty-eight cases occurred within our health system in Southern California, the largest number of cases in the United States to date. We aim to summarize the clinical features, diagnosis, treatments, and outcomes of these cases.MethodsWe reviewed the electronic health records of 28 patients with identified MC infection who had index coronary artery bypass (CABG) and/or valve replacement surgery between 2014 and 2016. All diagnoses were confirmed by cultures speciated to MC by 16S partial DNA sequencing or Karius testing, except for one case. Patients were grouped by clinical presentation of disseminated disease (n = 18) or localized disease (n = 10). Treatment delay was calculated from the time of initial presentation to the start date of antibiotics and evaluation for surgical intervention.ResultsAll patients who underwent CABG alone (n = 5) developed localized sternal wound infections, whereas patients who had valve replacement surgery (n = 23) developed either localized or disseminated disease. Disseminated disease carried a mortality rate of 40% in those with surgical source control vs. 72% in patients who were not surgical candidates (OR 6.6, 95% CI 0.8–55). The mortality rate of patients with localized sternal wound infections was 11% after incision/drainage and sternal wire removal (n = 9). Delay of antimicrobial treatment greater than 6 months in all 28 patients was associated with a mortality rate of 54% compared with 35% in patients who started treatment within 6 months (OR 2.2, 95% CI 0.47–10.35). Overall mortality rate of patients with MC infection was 42%.ConclusionDisseminated MC infection should be considered early in at-risk patients presenting with constitutional symptoms. In this review of 27 confirmed and 1 probable case of MC infection, disseminated infection only occurred in patients who underwent valve surgery while localized disease occurred in patients who underwent CABG. Surgical source control with early initiation of antimicrobial therapy is associated with improved outcomes. Optimal duration of antimicrobial treatment is still unknown.Disclosures All authors: No reported disclosures.

Identification of Fusarium spp. Associated with Potato Tubers in Upper Egypt by Morphological and Molecular Characters

Potato (Solanum tuberosum L.) is an important crop which holds promise for food security considering the global population growth rate. Fusarium dry rot is one of the most significant diseases of potato. To build up strategies for the control of this disease, it ought to be made primarily a correct diagnosis and identification of the pathogen. A total of 504 Fusarium isolates were recovered from potato tubers collected from Upper Egypt. Fusarium isolates were identified based on morphology and partial DNA sequencing of β-tubulin (TUB) genes. 62.5% of the isolates were identified as F. sambucinum, followed F. oxysporum (57.5%), then F. verticillioides (56.25%) and F. incarnatum (47.5%). All the tested Fusarium species were able to produce amylase. The pathogenicity of the isolates was tested by inoculation of healthy potato tubers; all of the tested isolates were pathogenic to healthy potato tubers. F. sambucinum had a highly virulent effect.

Molecular detection of Ehrlichia canis in Rhipicephalus sanguineus (s.l.) ticks in dogs and their domestic environment in Cuiaba, MT, Brazil

The central region of Brazil is known to be an endemic area for canine ehrlichiosis. Therefore, this study aims to determine the prevalence rates of E. canis infection in dogs and in Rhipicephalus sanguineus ticks collected from the dogs and their home environments. Serum samples and genomic DNA from the blood of 20 dogs and 299 ticks were analyzed by IFA and PCR assays in order to detect Ehrlichia canis antibodies and DNA. Nine (45%) of the 20 dogs were seropositive for E. canis, with titers ranging from 80 to 10240, and 6 dogs (30%) were positive for Ehrlichia spp. by PCR. Five free-living ticks were positive (2.89%, 95% confidence interval: 0.94-6.62%), as were six ticks attached to dogs (4.76%; 95% CI: 1.77-10.0%). The two groups showed a similar infection rate (P=0.395). Partial dsb DNA sequences of two samples from ticks were identical to each other and 100% (350/350 nucleotides) were identical to E. canis. Despite the high serological and molecular rates of canine ehrlichiosis in Cuiabá, the prevalence among infected ticks was lower than that found among dogs. However, adult ticks may remain infective much longer to ensure their infestation and infection of susceptible dogs.

Isospora svecica sp. n. (Apicomplexa: Eimeriidae), a new species of coccidium from the white-spotted bluethroat Luscinia svecica cyanecula (Aves: Passeriformes: Muscicapidae).

Using a combination of morphological and molecular data, we describe a new apicomplexan parasite, Isospora svecica sp. n., from the white-spotted bluethroat, Luscinia svecica cyanecula, from the Czech Republic. Oocysts were found in its intestinal tract. Sporulation was exogenous and took 1-3days. The oocysts were slightly ellipsoidal, of average size 26.17 × 20.33μm, with a smooth bilayered wall. Micropyle, oocyst residuum, and polar granules were absent. Sporocysts were bottle-shaped, of an average size of 18.82 × 8.82μm, with a thin, colourless wall. A conspicuous knob-like Stieda body was present. Substieda body was barely visible. Sporocyst residuum was present in the form of granules of various sizes. Sporozoites were banana-shaped and contained large anterior and small posterior refractile bodies. Partial DNA sequences of three genes were obtained from oocysts of Isospora svecica sp. n., being most closely related to other isosporans described from passerines. Little is known about the parasites of the avian family Muscicapidae, including coccidia, a highly prevalent parasitic protist group in all vertebrate classes. Only six species of the genus Isospora have so far been described in Muscicapidae, together with several "Isospora sp." that in fact most likely represent Isospora lacazei. The newly described Isospora svecica sp. n. differs morphologically from other coccidia reported from muscicapid birds, and represents the first coccidian species described from Luscinia svecica.

Isolation of Partial Housekeeping Genes on Tuntun Angin (Elaeocarpus floribundu)

Some genes like 18S rRNA, 26S rRNA, elongation factor 1-alpha (EF1a), and beta-tubulin (TUB) are members of housekeeping genes group that are commonly used as internal control in gene expression study. This study aimed to isolate those four housekeeping genes of tuntun angin (Elaeocarpus floribundus). The research material included fresh leaves of E. floribundus that were picked up from Kajuik Lake in Riau Province and four primer pairs. The procedures consisted of total DNA isolation using Genomic DNA Mini Kit Plant (Geneaid), polymerase chain reaction (PCR), electrophoresis on 1% agarose gel, sequencing, and bioinformatic analysis. This study has been isolated 18S rRNA, 26S rRNA, EF1a, and TUB genes with the size of 422 bp, 922 bp, 856 bp, and 877 bp, respectively. The EF1a and TUB genes has never been reported in Elaeocarpaceae family. Thus, those partial DNA sequences are the first sequences reported from this species and can be used as reference genes in this plant after validation.