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DNA barcoding based on the cytochrome oxidase I gene is increasingly used in black flies (Diptera: Simuliidae), but extensive data for larger areas are still rare. Slovakia, with well-explored black fly fauna, was chosen to verify the reliability of DNA barcoding for species identification. The DNA barcoding region of the COI gene of 235 individuals of 25 black fly species from Slovakia was sequenced. Among them, 30 sequence clusters with assigned Barcode Index Numbers (BINs) were identified, and 5 of them were recorded for the first time. The average intraspecific genetic divergence was 0–3.24%, whereas the average interspecific divergence was 12.3–17.8%. Based on the barcode sequence, 14 species could be identified unambiguously, and 3 of them (Prosimulium latimucro, Simulium costatum, S. degrangei) are split into two or more barcode clusters. In eleven species, some degree of barcode sharing occurred, often combined with barcode splitting. The results showed hidden diversity as well as cases of shared barcode sequences among the studied species. Further investigation using other molecular markers is necessary due to the overlap of intraspecific and interspecific variability.

There is little fossil evidence for cave arthropods. Small size, disfunctional wings, extremely long antenna, dense minute setation, palidity and reduced eyes are typical adaptations to life in caves shown by the cockroach Mulleriblattina bowangi gen. et sp. n., found together with epigeic Crenocticola svadba sp. n., both from Myanmar amber and belonging to the cavernicolous cockroach family Nocticolidae. These lineages of earliest still living cavernicoles suggest large, numerous caverns, lava tubes or caves within the source area. They provide the first unequivocal evidence for the Mesozoic origin of any living troglomorphic organism, and explain the “long branches” in DNA analyses. Phylogenetic trees show little hierarchical structure and place Latindiinae and Myrmecoblatta within the explosively radiating Nocticolidae. Biogeography indicates a common cosmopolitan Early Cretaceous ancestor except for 8 (of 49) species of true Nocticolidae, which diverged during the Late Cretaceous breakup of Gondwana. A review of all troglofauna documents no other unequivocal pre-Cenozoic cave biotas (including vertebrates). Stable environments and small populations result in a short-time (˂3 Ma) origination of bizarre forms and long term extinctions (˃30 Ma).

Direct fossil evidence for Mesozoic flower pollination is scarce. Umenocoleoid micro-cockroaches Lepidopterix vegrandis gen. et sp. n. (Lebanese amber) and Antophiloblatta hispida gen. et sp. n. (Myanmar amber) possess size, form, complex coloration pattern and surface structure, cryptic with potentially entomophilous angiosperms Tropidogyne pentaptera Poinar, 2017 and Antiquifloris latifibris Poinar et Buckley, 2016 flower petals and sepals. Putative pollen grains attached to the latter adult indicates pollination, while reduced mouthparts suggest fluid nectar feeding. Spongistoma angusta gen. et sp. n. (Myanmar amber) has a narrow body and mandibles nearly entirely reduced with a unique “proboscis” forming sponging/ sucking mouthparts. In addition putative Classopolis Pflug, 1953 gymnosperm pollen is attached to adults and immature individuals of Vzrkadlenie miso gen. et sp. n. (Myanmar amber). Together with possible angiosperm pollination by Formicamendax vrsanskyi Hinkelman, 2019 and cycas pollination by immature individuals of alienopterid larvae, the evidence for early cockroach pollination is now substantial. Additionally unique is the forewing surface of L. vegrandis with photonic crystal structures within the scales.

The development of CD4+ T helper cells is determined by the set of transcription factors and the genes these transcription factors transcribe. In this review, we describe the basic nature of Th1, Th2, Th9, Th17, T-follicular helper (Tfh), gamma delta (γδ) T cells, and T-regulatory (Treg) cells subsets, their master regulator transcription factors and their corresponding signature cytokine production profiles. Cellular immunity plays important role during virus infection. Optimal immune response to viral infections require a gentle balance of effector responses to clear the infected cells and regulatory mechanism to prevent immunopathology. The behavior of the helper cells differs with each virus - while in some cases, the response is beneficial; in other cases, it is harmful. We discuss the protective and pathological role of T cell immunity against influenza A virus (IAV), respiratory syncytial virus (RSV), immunodeficiency virus type 1 (HIV-1), and hepatitis B virus (HBV) infection. Keywords: T cell; cytokine; influenza virus; respiratory syncytial virus; hepatitis B virus; human immunodeficiency virus type 1.

To establish infection, a wide variety of pathogens, including viruses, have evolved a number of strategies to avoid immune elimination. Viruses have acquired and optimized molecules that interact with the host chemokine network in order to disrupt immune surveillance and defense of vertebrates, helping to promote cell entry, facilitating dissemination of infected cells, and evasion the immune response. Viral immunomodulators include ligands, chemokine receptors and chemokine-binding proteins (vCKBPs) functioning as either cell surface receptor mimics, ligand mimics, or secreted chemokine-binding proteins. vCKBPs specifically modulate chemokine gradient formation and ligand-receptor recognition when they have a potential to even completely block chemokine-mediated responses to viral infection. Members of only two virus families (Herpesviridae and Poxviridae) encode vCKBPs capable of sequestering host chemokines through either the chemokine receptor, GAG-binding pocket, or both, which may result in the inhibition of chemotaxis in vivo. Here, we focused on vCKBPs encoded by α-, β-, and γ-herpesviruses, of which several have been experimentally used as anti-inflammatory or anti-immune reagents in animal models. Current results suggest that vCKBPs could be used to regulate the activity of both chemokines and chemokine receptors for the treatment of infections such as AIDS, diseases such as arthritis, neurotrauma, inflammatory CNS disorders, atherosclerosis, transplant rejection, and metastatic spread and angiogenesis. Better understanding of vCKBPs biology will help evaluate, which human diseases related to chemokine network dysregulation might be effectively treated with these novel promising immunomodulatory drugs to enable the manipulation of chemokine functions and leukocyte trafficking. Keywords: herpesviruses; chemokine-binding proteins; chemokines; immunomodulation viral infection, chemokines and viral immunomodulators.

Jejunal diverticulosis is a rare diagnosis that occurs mainly in old age, more often in men than in women. It is usually an incidental diagnosis of unclear aethtiology. In some cases, visceral myopathy can also be the cause. It is most often manifested by abdominal pain and bleeding. Bleeding from the small intestinal diverticula represents only 0.6-5% of all small intestinal bleeding.The authors describe the case of a 66-year-old man with massive gastrointestinal bleeding who did not respond to conservative hemostyptic treatment. Following negative gastrofibroscopic and colonoscopic examinations, an angioCT examination was indicated, which revealed a source of bleeding in the jejunal diverticula. The patient was indicated for surgical treatment. The extent of bleeding was determined by perioperative enteroscopy and subsequently, the affected jejunal segment was segmentally resected with a primary anastomosis.Bleeding from the jejunal diverticula is a very rare diagnosis, which poses challenges in the diagnostic process in particular. Capsule enteroscopy plays an important role in the diagnosis, as well as CT angiography and scintigraphy in the event of massive bleeding. In addition to conservative treatment, the embolization of a bleeding vessel may subsequently be used in therapy. In indicated cases, surgical resection treatment is also possible.

Recently, non-cellular viral agents became the focus of a large number of scientific groups. A prominent and widespread group of these viruses are flaviviruses, which include, for example, Zika virus, Dengue fever virus, tick-borne encephalitis virus and West Nile virus. There is a considerable diversity among these viruses, however, highly conserved proteins can be found throughout this viral genus. The largest and most conserved protein encoded by flaviviruses is the nonstructural NS5 protein. Its N-terminal domain bears the methyltransferase (MTase) activity. Thanks to the methylation of its genome, it allows the virus to initiate translation and at the same time mask it from the host's immune system. By blocking the active site of this enzyme with a small molecule, viral infection could be stopped not only in one flavivirus, but, due to the high conservation of MTases, in all other flaviviruses. This diploma thesis deals with the aforementioned MTase domain of the NS5 protein, specifically of the West Nile virus (WNV). After designing an insert encoding the WNV MTase domain, amplifying it and ligating it into the vector, the MTase domain was prepared by a recombinant expression, followed by purification. Subsequently, complexes of the protein with small molecules (MTase ligands) were formed, in...

Radiation-associated angiosarcoma (RAAS) is a rare and serious complication of breast irradiation. Due to the rarity of the condition, clinical experience is limited and publications on this topic include only retrospective studies or case reports.All patients diagnosed with RAAS between January 2000 and December 2017 in twelve centers across the Czech Republic and Slovakia were evaluated.Data of 53 patients were analyzed. The median age at diagnosis was 72 (range 44-89) years. The median latency period between irradiation and diagnosis of RAAS was 78 (range 36-172) months. The median radiation dose was 57.6 (range 34-66) Gy. The whole breast radiation therapy with radiation boost to the tumor bed was the most common radiotherapy regimen. Total mastectomy due to RAAS was performed in 43 patients (81%), radical excision in 8 (15%); 2 patients were not surgically treated due to unresectable disease. Adjuvant chemotherapy followed surgical therapy of RAAS in 18 patients, 3 patients underwent adjuvant radiotherapy. The local recurrence rate of RAAS was 43% and the median time from surgery to the onset of recurrence was 7.5 months (range 3-66 months). The 3-year survival rate was 56%, the 5-year survival rate was only 33%. 46% of patients died during the follow-up period.The present data demonstrate that RAAS is a rare condition with high local recurrence rate (43%) and mortality (the 5-year survival rate was 33%.). Early diagnosis of RAAS based on biopsy is crucial for treatment with radical intent. Surgery with negative margins constitutes the most important part of the therapy; the role of adjuvant chemotherapy and radiotherapy is still unclear.

Viruses have evolved sophisticated strategies to subvert immunity to benefit overall viral fitness. Human cytomegalovirus (HCMV, β-herpesvirus) represents a paradigm of very effective hijacking of gene functions that imitate host encoded immunomodulatory proteins. This co-evolution with the host immune system allowed for establishment of lifelong persistence. The HCMV infection is largely asymptomatic in healthy persons; however, it can induce serious disease in immunocompromised individuals. For this reason, great attention is paid to the development of therapeutics based on HCMV immunomodulatory 'tricks' as well as to the search for active vaccine against HCMV. While comparing the HCMV clinical isolates with extensively passaged laboratory strains, the unique long (UL) b' locus was commonly found to be deleted in HCMV genome while adapted to replication in human fibroblasts in vitro. This missing region, called UL/b' region, encodes up to 22 canonical genes with different functions, as of targeting cellular tropism (e.g. UL133-UL138); viral entry and assembly (e.g. UL128, UL130, UL131A); regulation of immunological synapses (e.g. UL135); inhibition of NK and T cell function (e.g. UL141, UL142, UL148, UL144), ablating activity (e.g. UL146, UL147), but mainly aimed at manipulating the host immune response. Moreover, the presence of UL/b' genomic region dramatically correlates with adverse effects in vaccinated persons, indicating that viral genes in this region play a significant role in controlling virulence. Here, we review how HCMV shapes our immunity by hijacked genes originated from UL/b' locus, discuss their impact in immunomodulation mechanism and how this knowledge may translate to clinical applications. Keywords: immunomodulation; HCMV genes; UL/b' locus; NK cell function; HCMV vaccine; immunity; immunotherapeutics.

Infection of human MRC-5 cells and mouse NIH-3T3 cells with a murine gamma-herpesvirus (MuHV-4 strain 68; MHV-68) photoinactivated by visible light in the presence of methylene blue (MB) resulted in nonproductive infection and the appearance of morphologically transformed cells. Two stably transformed cell lines were derived from both of these cell types and were confirmed to contain both viral DNA and antigen. Next, a quiescent MHV-68 infection in MRC-5 and NIH-3T3 cells was established after cultivation at 41°C in the presence of phosphonoacetic acid. Following the exposure of quiescently infected cells to visible light for 120 s (5 times daily for 6 days) in the presence of MB, both MRC-5 and NIH-3T3 cells were observed to acquire transformed phenotypes. The cytopathic effect was observed in cells after 4-5 passages, after which the cells degenerated. However, when human interferon (IFN)-α and mouse IFN-β were added to the media of quiescently infected MRC-5 and NIH-3T3 cells during the photoinactivating procedure, 2 stable transformed cell lines containing both viral DNA and the antigen were obtained and resembled those attained following nonproductive infection with photoinactivated virus.

A collaborative European DNA Profiling (EDNAP) Group exercise was undertaken to assess the performance of an earlier described SNaPshot™-based screening assay (denoted mini-mtSNaPshot) (Weiler et al., 2016) [1] that targets 18 single nucleotide polymorphism (SNP) positions in the mitochondrial (mt) DNA control region and allows for discrimination of major European mtDNA haplogroups. Besides the organising laboratory, 14 forensic genetics laboratories were involved in the analysis of 13 samples, which were centrally prepared and thoroughly tested prior to shipment. The samples had a variable complexity and comprised straightforward single-source samples, samples with dropout or altered peak sizing, a point heteroplasmy and two-component mixtures resulting in one to five bi-allelic calls. The overall success rate in obtaining useful results was high (97.6%) given that some of the participating laboratories had no previous experience with the typing technology and/or mtDNA analysis. The majority of the participants proceeded to haplotype inference to assess the feasibility of assigning a haplogroup and checking phylogenetic consistency when only 18 SNPs are typed. To mimic casework procedures, the participants compared the SNP typing data of all 13 samples to a set of eight mtDNA reference profiles that were described according to standard nomenclature (Parson et al., 2014) [2], and indicated whether these references matched each sample or not. Incorrect scorings were obtained for 2% of the comparisons and derived from a subset of the participants, indicating a need for training and guidelines regarding mini-mtSNaPshot data interpretation.

The M3 protein (M3) encoded by murine gammaherpesvirus 68 (MHV-68) is a unique viral immunomodulator with a high-affinity for a broad spectrum of chemokines, key mediators responsible for the migration of immune cells to sites of inflammation. M3 is currently being studied as a very attractive and desirable tool for blocking the chemokine signaling involved in some inflammatory diseases and cancers. In this study, we elucidated the role of M3 residues E70 and T272 in binding to chemokines by examining the effects of the E70A and T272G mutations on the ability of recombinant M3, prepared in Escherichia coli cells, to bind the human chemokines CCL5 and CXCL8. We found that the E70A mutation enhanced binding of M3 to CCL5 two-fold but had little effect on its binding to CXCL8. In contrast, the T272G mutation was found to be important for the thermal stability of M3 and significantly decreased M3's binding to both CCL5 (by about 4×) and CXCL8 (by about 5×). We also constructed in silico models of the wild-type M3-CCL5 and M3-CCL8 complexes and found substantial differences in their physical and chemical properties. M3 models with single mutation E70A and T272G suggested the role of E70 and T272 in binding M3 protein to chemokines. In sum, we have confirmed that site-directed mutagenesis could be an effective tool for modulating the blockade of particular chemokines by M3, as desired in therapeutic treatments for severe inflammatory illnesses arising from chemokine network dysregulation.

TEAD proteins belong to a significant family of transcription factors that contribute to the regulation of organism growth and cell differentiation during its development by activating the expression of a wide variety of genes. This family shares two highly conserved sites, the TEA DNA binding domain, after which the proteins have been named, and the domain by which transcription factors bind other coactivators. Because TEAD proteins are not able to activate transcription themselves, they interact with a number of coactivators. These coactivators allow the transcription of the gene of interest to be regulated. Failure of TEAD protein activity regulation can lead to cancer. Therefore, TEAD family proteins nowadays play an important role in the development of new anticancer drugs. One way of inhibiting these proteins is to block the active site in their DNA binding domain, thus, to block their binding to DNA. This bachelor thesis deals with recombinant expression of said DNA binding domain of transcription factor TEAD1, which is extended by amino acids in unstructured regions. After finding suitable conditions of protein production, we proceeded to large volume production which was followed by purification and protein identity verification. Finally, the ability of the produced protein to interact...

Murine gammaherpesvirus 68 (MHV-68) is a natural pathogen that infects murid rodents which serve as hosts for Dermacentor reticulatus and Ixodes ricinus ticks. For the first time, MHV-68 was detected in immature I. ricinus ticks feeding on lizards trapped in Slovakia. Later on, MHV-68 infection was detected in D. reticulatus and Haemaphysalis concinna ticks collected on vegetation, which supported the idea that ticks can acquire the virus from feeding on infected hosts. Here, we report MHV-68 infection, which was detected by nested PCR, in D. reticulatus and I. ricinus adult ticks and I. ricinus nymphs collected in five geographically isolated localities, in west, southwest, south and central Slovakia. Viral incidence in ticks was 46.7% (121/259) without considering the season, site of collection and tick species and their life stage. MHV-68 infection was detected in all five localities investigated and in both tick species. Here, for the first time, we report MHV-68 infection in I. ricinus nymphs collected from the vegetation. The finding of virus in ticks from five separated localities suggested that ticks became infected with MHV-68 via feeding on infected rodents; thus, this virus might be a newfound natural pathogen in ticks.

The black fly Prosimulium italicum Rivosecchi, distributed in the Apennines and Sicily, was described as a subspecies of Prosimulium hirtipes (Fries), based on a few morphological details. It subsequently was considered conspecific with P. hirtipes and the name was synonymized. Analyses of polytene chromosome banding patterns and sequences of mitochondrial DNA (COI and COII) revealed deep genetic divergence between P. italicum from Italy and P. hirtipes from northern and central Europe and confirmed the species status of P. italicum . Populations of P. italicum either lack chromosomal inversion IS-9 or carry it as an X-chromosome polymorphism, whereas all analyzed populations of P. hirtipes (Slovakia, Sweden, England, and Scotland) are fixed for IS-9 . The average K2P genetic distance was 3.7% between P. italicum and P. hirtipes from northern Europe (Sweden) and 4.3 % between P. italicum and P. hirtipes from central Europe (Slovakia). Cytogenetic analysis showed the presence of two cytoforms of P. hirtipes (‘A’ in Sweden and Slovakia and ‘B’ in England and Scotland) and two cytoforms of P. italicum (‘A’ in Sicily and ‘B’ in Campania and Basilicata), all of which differ in their sex chromosomes and autosomal polymorphisms, suggesting that P. hirtipes and P. italicum might each be a complex of cryptic species.

Kúdela, Matúš, Adler, Peter H., Kúdelová, Tatiana (2018): Taxonomic status of the black fly Prosimulium italicum Rivosecchi (Diptera: Simuliidae) based on genetic evidence. Zootaxa 4377 (2): 280-290, DOI: https://doi.org/10.11646/zootaxa.4377.2.8

Human dermal fibroblasts and mouse NIH/3T3 cells acquired the transformed phenotype (‘criss-cross' pattern of growth) after infection with ultraviolet-irradiated murine gammaherpesvirus (MuHV-4 strain 68; MHV-68). These cells with changed phenotype could be serially cultured for 5-6 passages (35-40 days), and then they entered into crisis and most of them died. In a small number of cultures, however, foci of newly transformed cells appeared from which two stable cell lines were derived. After 6-9 cell culture passages of the MHV-68 transformed cell lines, MHV-68 DNA and virus antigen could be detected by PCR and immunofluorescence assay along with the disappearance of actin bundles, indicating that both transformed cell lines might be oncogenic.

M3 protein of murine gammaherpesvirus 68 (MHV-68) was identified as a viral chemokine-binding protein 3 (vCKBP-3) capable to bind a broad spectrum of chemokines and their receptors. During both acute and latent infection MHV-68 M3 protein provides a selective advantage for the virus by inhibiting the antiviral and inflammatory response. A unique mutation Asp307Gly was identified in the M3 protein of murine gammaherpesvirus 72 (MHV-72), localized near chemokine-binding domain. Study on chemokine-binding properties of MHV-72 M3 protein purified from medium of infected cells implied reduced binding to some chemokines when compared to MHV-68 M3 protein. It was suggested that the mutation in the M3 protein might be involved in the attenuation of immune response to infection with MHV-72. Recently, Escherichia coli cells were used to prepare native recombinant M3 proteins of murine gammaherpesviruses 68 and 72 (Pančík et al., 2013). In this study, we assessed the chemokine-binding properties of three M3 proteins prepared in E. coli Rosetta-gami 2 (DE3) cells, the full length M3 protein of both MHV-68 and MHV-72 and MHV-68 M3 protein truncated in the signal sequence (the first 24 aa). They all displayed binding activity to human chemokines CCL5 (RANTES), CXCL8 (IL-8), and CCL3 (MIP-1α). The truncated MHV-68 M3 protein had more than twenty times reduced binding activity to CCL5, but only about five and three times reduced binding to CXCL8 and CCL3 when compared to its full length counterpart. Binding of the full length MHV-72 M3 protein to all chemokines was reduced when compared to MHV-68 M3 protein. Its binding to CCL5 and CCL3 was reduced over ten and seven times. However, its binding to CXCL8 was only slightly reduced (64.8 vs 91.8%). These data implied the significance of the signal sequence and also of a single mutation (at aa 307) for efficient M3 protein binding to some chemokines.

Gammaherpesviruses-encoded replication and transcription activator (Rta) (ORF50) plays an essential role in the initiation of viral lytic gene expression and reactivation from latency. The Rta expression is influenced by many viral and cellular factors, including epigenetic modifications, mainly DNA methylation and histone modifications. Murine gammaherpesvirus 68 (MHV-68), belonging to the species Murid herpesvirus (MuHV-4), is widely used as a model to study human gammaherpesvirus infections in vitro as well as in vivo. Recent studies of the MHV-68 Rta promoter revealed the effect of DNA demethylation and histone acetylation, induced by the inhibitor of histone deacetylase trichostatin A (TSA), on the MHV-68 reactivation from latency. Two other strains of MuHV-4, MHV-72 and MHV-4556, possess several unique properties, which distinguish them from strain MHV-68. Recently discovered reduced capacity of MHV-72 and MHV-4556 to reactivate from latency may be related to different methylation/demethylation patterns of the promoter regulatory region of the Rta. Here, we investigated the epigenetic regulation of the Rta promoter of three murine gammaherpesvirus strains, MHV-68, MHV-72 and MHV-4556, during latency and reactivation in vivo. However, we did not find any differences between Rta of MHV-68, MHV-72 and MHV-4556 and its epigenetic regulation during lytic infection, latency and de novo infection after ex vivo and in vivo reactivation induced by TSA. We confirmed that the treatment with TSA successfully induced demethylation of the Rta promoter regions of all three studied strains. Moreover, we have shown that the primary sequence of Rta and its promoter is identical for all three strains.

The new, small cavernicolous species Helmablatta louisrothi gen. et sp. n. (Nocticolidae) from the Tan-Phu cave (Vietnam) is one of the most morphologically interesting cockroaches. The extremely modified upstanding tergal gland composite from three tergites and may serve for gripping the female head during copulation. This presumption is supported by the presence of a central big hook on tergite 8. Furthermore, both wing pairs are uncommonly adapted to help releasing sex pheromones without raising the wings. Histone 3 DNA-based maximum likelihood analyses indicate a recent origin and close phylogenetic relationship between Nocticola spp. and Helmablatta sp.—consistent with the Quaternary age of the source lava tubes.

Helmablatta louisrothi Vidlička, Vr��ansk�� et Deharveng, sp. n. Type material. Holotype ���male (Vn06 253): southeastern region of Vietnam: T��n Ph�� area, Dinh Quan district of Đ���ng Nai, Lava tube cave (11��15'1.72''N, 107��24'14.37''E), altitude 180 m; collected by hand near bat guano, leg. Louis Deharveng & Anne Bedos; coll. Museum National d'Histoire Naturelle, Paris, France. Data deposition in GenBank, under Nocticolidae gen. n.���KR814821. Description. Male. Body pale, brownish yellow, nearly translucent, only apex of maxillae darker. Head very big (one third of body length), ratio of head length/width = 1.45 (1.167/ 0.805 mm), oval, invisible from above (opisthognathous at rest); clypeus and labrum short, overlaps mandible only partly; mandibular tip with strongly sclerotized teeth; eyes small (0.15 mm long)���about 40-faceted, oblong, narrow, strongly reduced, facets marginally obvious; lateral and central ocelli absent. Maxillary palps very long (length of palpomeres 1st���5th: 0.096, 0.172, 0.310, 0.316, 0.340 mm), first and second palpomeres very short, 3th to 5th palpomeres approximately equal, apex of last segment obliquely truncated. Labial palps three-segmented (length of palpomeres 1st���3rd: 0.191, 0.220, 0.306 mm), last segment rounded on apex. Antennae longer than body (preserved part 5.1 mm long, scapus 0.145/ 0.085 mm), slender, complete number of antennomeres is unknown, only 47 segments remained, scape and pedicel moderate elongated, following 10 segments as long as wide, other segments two times longer than wide. Pronotum parabolic, lateral hind corner slightly oblong (width/length = 1.360/ 0.964 mm). Tegmina do not touch, shortened (1.944 mm long), reaching to posterior margin of 6th abdominal segment, in apical third strongly tapered (width in middle 0.655 and in apical third 0.361 mm) and sinuous on inner side, uncovering (outflanking) the tergal gland, apex volute internal, venation invisible with the exception of indistinct Sc vein, upper surface and inner margin of sinuous part covered by setae, basis with a few spines (Figs. 2 e, 3e). Hind wings strongly reduced (0.990/ 0.428 mm), widest in apical third, without visible venation, with unique very deep incision in apical part, upper surface with very sparse and small setae, basis with a few spines (Fig. 3 f). All legs, in proportion to body, very large. Procoxae elongated, tapered, meso- and metacoxae triangular, robust, proximally with a few small spines; femora on anterior side covered with small spines, genicular spines of femora present, length of fore-, mid- and hind femora 1.214, 1.275 and 1.579 mm, respectively; anteroventral margin of fore femur with alternate large and small spines irregularly disposed, and a large distal spine (intermediate Type A1 ��� B1���terminology follow Roth 2003); tibia with several small spines, distal end with four large spines, length of fore-, mid- and hind tibia 0.801, 1.020 and 1.603 mm, respectively; tarsomeres with small spines in three rows (Figs. 3 a, b, c); tarsomere 1 ��� 5 length: (front-, mid- and hind-leg): (0.607, 0.777, 0.923 mm), (0.218, 0.231, 0.316), (0.085, 0.121, 0.151), (0.073, 0.085, 0.090), (0.170, 0.170, 0.180); tarsal claws simple, symmetrical; pulvilli and arolia absent; basitarsi of all legs longer than the four other tarsomeres combined. Abdomen very short (1.756/ 1.090 mm) with very distinct tergal gland. Abdominal terga T3���T5 strongly specialized (Figs. 1a, 2a, b, c, 3d, 4a, b, c), together form very sophisticated and unique tergal gland equipment, where T4 is extremely extruded over surface of abdomen; abdominal tergite T8 create in the middle big anteriorly incurved hook (���nipper���)���extremely rigid part of the insect, interfere with cavity on T7; cerci (incomplete) with very long segments in posterior part and long sensilla on the apex, surface covered with setae (Figs. 1a, b). Supra-anal plate wide, subtrapezoidal, with central concave hind margin (Figs. 1a, d, 2a, 4a, b, d). Sub-genital plate symmetrical, at the sides with two similar very short styles, inter-stylar margin convexly rounded (Fig. 1b). Genital hook (L3) projects (0.388 mm long) on the left side, subapicali wide (0.062 mm) with several strong setae in inner margin, the apex of hook strongly narrow (0.021 mm), long (0.128 mm) and rectangular curved (Figs. 4 b, d). Etymology. New cockroach species is named in honour of Louis M. Roth (*1918 ��� ���2013), one of the greatest ���cockroachologist��� of the 20th century who published the latest revision of Nocticolidae., Published as part of Vidli��ka, ��ubom��r, Vr��ansk��, Peter, K��delov��, Tatiana, K��dela, Mat����, Deharveng, Louis & Hain, Miroslav, 2017, New genus and species of cavernicolous cockroach (Blattaria, Nocticolidae) from Vietnam, pp. 361-375 in Zootaxa 4232 (3) on pages 365-370, DOI: 10.11646/zootaxa.4232.3.5, http://zenodo.org/record/312401, {"references":["Roth, L. M. (2003) Systematics and phylogeny of cockroaches (Dictyoptera: Blattaria). Oriental Insects, 37, 1 - 186. https: // doi. org / 10.1080 / 00305316.2003.10417344"]}

Herpesviruses are a large group of DNA viruses infecting mainly vertebrates. Murine gammaherpesvirus 68 (MHV68) is often used as a model in studies of the pathogenesis of clinically important human gammaherpesviruses such as Epstein-Barr virus and Kaposi's sarcoma-associated herpesvirus. This rodent virus appears to be geographically widespread; however, its natural transmission cycle is unknown. Following detection of MHV68 in field-collected ticks, including isolation of the virus from tick salivary glands and ovaries, we investigated whether MHV68 is a tick-borne virus. Uninfected Ixodes ricinus ticks were shown to acquire the virus by feeding on experimentally infected laboratory mice. The virus survived tick molting, and the molted ticks transmitted the virus to uninfected laboratory mice on which they subsequently fed. MHV68 was isolated from the tick salivary glands, consistent with transmission via tick saliva. The virus survived in ticks without loss of infectivity for at least 120 days, and subsequently was transmitted vertically from one tick generation to the next, surviving more than 500 days. Furthermore, the F1 generation (derived from F0 infected females) transmitted MHV68 to uninfected mice on which they fed, with MHV68 M3 gene transcripts detected in blood, lung, and spleen tissue of mice on which F1 nymphs and F1 adults engorged. These experimental data fulfill the transmission criteria that define an arthropod-borne virus (arbovirus), the largest biological group of viruses. Currently, African swine fever virus (ASFV) is the only DNA virus recognized as an arbovirus. Like ASFV, MHV68 showed evidence of pathogenesis in ticks. Previous studies have reported MHV68 in free-living ticks and in mammals commonly infested with I. ricinus, and neutralizing antibodies to MHV68 have been detected in large mammals (e.g., deer) including humans. Further studies are needed to determine if these reports are the result of tick-borne transmission of MHV68 in nature, and whether humans are at risk of infection.

Three strains of herpes simplex virus, K17syn- and HSZPsyn+ of type 1 (HSV-1) and USsyn- of type 2 (HSV-2), were photoinactivated in the presence of methylene blue and used to infect 3 cell lines, normal human lung tissue cells (MRC-5), mouse epithelial cells (NIH3T3), and human lung carcinoma cells (A549). The virus titer and phenotype of cells were evaluated to compare the characteristics of normal and carcinoma cells infected with non-syncytial (non-syn) and syncytial (syn) strains of herpes simplex viruses. We found that the cells of both normal cell lines infected with photoinactivated K17syn- and USsyn- but not HSZPsyn+ acquired transformed phenotype accompanied by the presence of virus. Surprisingly, the infection with photoinactivated viruses K17syn- and USsyn- but not HSZPsyn+ resulted in the suppression of the transformed phenotype of A549 cells. Using nested PCR, herpesviral DNA was identified in newly transformed cells and cells that lost the transformed phenotype. The effect of putative herpesvirus-related growth factors (HRGF) produced by cells infected with photoinactivated viruses was quantified and compared. Since methylene blue is currently used in phototherapy of herpetic lesions, these results raise the question of whether such therapy is risky to human health.

Murine herpesvirus 68 (MHV-68) is a natural pathogen that infects murid rodents, which serves as hosts for Ixodes ricinus ticks. For the first time, MHV-68 was detected in immature I. ricinus ticks feeding on Lacerta viridis lizards trapped in Slovakia, which supports the idea that ticks can acquire the virus from feeding on infected hosts. The recent discovery of MHV-68 infection and MHV-68 M3 gene transcripts in Dermacentor reticulatus ticks collected in Slovakia also supports this suggestion. Here, for the first time, we report MHV-68 infection, which was detected by nested PCR, in I. ricinus adults collected from the vegetation, and the viral load in infected ticks was determined by quantitative PCR. The viral incidence in ticks was 38.1% (21/55), and the viral load varied from 1.5 × 103 to 2.85 × 104 genome copies per tick. These results suggest that the I. ricinus ticks became infected with MHV-68 from biting infected rodents; thus, I. ricinus ticks may play a role in the spread of this virus in nature.

Background: Murine gammaherpesvirus 68 is assumed to be a natural pathogen of murid rodents. Previous investigations of MHV68 in field-collected Dermacentor reticulatus, Ixodes ricinus, and Haemaphysalis concinna ticks support the idea that ticks acquire the virus from feeding on infected hosts. Based on our previous finding of a live MHV-68 capable to replicate in mammalian cells, we aimed to investigate if transcripts of MHV-68 are present in D. reticulatus ticks and to determine the amount of MHV-68 in these ticks. Methods: This study utilized a sensitive nested RT-PCR method to detect transcripts of the early-late M3 gene of MHV-68, then nested PCR to screen MHV-68 presence and real-time PCR to quantify virus infectious dose in ticks. Results: Transcripts of MHV-68 M3 gene were detected in 10 out of 11 questing ticks. MHV-68 was detected in 38 out of 48 questing ticks, in which an infectious dose of MHV-68 varies from 2.2 × 104–8.6 × 106 copies of the virus genome . Conclusion: We report the first evidence of MHV-68 transcription and infectious dose of MHV-68 in field collected D. reticulatus ticks. Results provide unique evidence that ticks could act as a reservoir of gammaherpesvirus, which could be capable of replication.

Summary. – This article is a brief summary of efforts to generate mutant herpesviruses for investigating and assigning gene functions of herpesviruses in replication and pathogenesis. While a full review of all herpesviruses is beyond the scope of this review, we focused our attention on the prototype of the herpesvirus subfamily – herpes simplex virus and murine gammaherpesvirus that serves as an excellent animal model to study human gammaherpesvirus pathogenesis. Furthermore, our present knowledge of essential, non-essential, and common genes of herpesviruses as well as of accessory genes that are currently being studied with the help of the bacterial artificial chromosome (BAC) system will also be discussed. This system facilitates the analysis of herpesviral genes with potential for use in gene therapy or as anti-cancer therapeutics.

Here we would like to provide a brief overview of the modern history of Marek΄s disease (MD) research with a focus on the most recent developments in experimental work and we will try to sum up their impact on the understanding of the biological properties of Marek΄s disease type 1 (MDV-1), the only representative of the Mardivirus genus causing fatal lymphoproliferative disease in poultry. We will also compare MDV-1 with other serologically-related poultry herpesviruses, Marek΄s disease virus type 2 (MDV-2) and herpesvirus of turkeys (HVT). Although MD was first described at the beginning of the last century, proper characterization of its biological impact on poultry production and utilization of molecular biology methods for detailed characterization of causative agent MDV-1 were introduced only in recent decades. However, many characteristics of MD infection, pathogenesis and vaccine protection mechanisms remain unclarified, though novel methods bring a challenge for better understanding of these unanswered questions.

Simulium (Simulium) argyreatum Meigen, 1838 New records. Upper reach of River Čabranka (19), larvae; upper reach of River Kupa (20), larvae., Published as part of Ivkovi��, Marija, K��dela, Matu�� & K��delov��, Tatiana, 2016, Blackflies (Diptera: Simuliidae) in Croatia: species richness, distribution and relationship to surrounding countries in Zootaxa 4109 (1), DOI: 10.11646/zootaxa.4109.1.2, http://zenodo.org/record/266198, {"references":["Meigen, J. W. (1838) Systematische Beschreibung der bekannten europaischen zweiflugeligen Insecten 7 (supplementary volume). Schulz, Hamm, Germany. 434 pp. + 8 plates."]}

Simulium (Boophthora) erythrocephalum (De Geer, 1776) Literature reference. River Sava near Zagreb (Baranov 1942) (7); River Kara��ica (Živković 1967 a, 1967 b, 1969, 1970 b) (11); River Drava at Osijek (Živković 1967 a, 1967 b, 1969, 1970 b) (12); River Danube at Batina (Živković 1967 a, 1967 b) (14); River Danube at Dalj (Živković 1967 b, 1971) (15); River Danube at Borovo (Živković 1967 b, 1971) (16); River Danube at Vukovar (Živković 1967 b, 1971) (17); River Danube at ��arengrad (Živković 1967 b) (18); Kopački Rit (Sudarić Bogojević et al. 2009) (13). New records. River Glina (28), larvae and pupae; River Krapina (4), larvae; River Ilova (9), larvae., Published as part of Ivkovi��, Marija, K��dela, Matu�� & K��delov��, Tatiana, 2016, Blackflies (Diptera: Simuliidae) in Croatia: species richness, distribution and relationship to surrounding countries in Zootaxa 4109 (1), DOI: 10.11646/zootaxa.4109.1.2, http://zenodo.org/record/266198, {"references":["De Geer, C. (1776) Memoires pour servir a l'histoire des insectes 6. Grefing, Stockholm, 523 pp.","Baranov, N. I. (1942) Boophthora argyreata Mg. U okolini Zagreba. Veterinarski Arhiv, 12, 209 - 2011.","Zivkovic, V. (1967 a) Invazija Simulium erythrocephalum De Geer, 1776 (Diptera, Simuliidae) u Vojvodini. Glas Srpske Akademije Nauka i umetnosti, odeljenje medicinskih nauka, 20, 163 - 170.","Zivkovic, V. (1967 b) Simulide (Diptera, Simuliidae) Dunava na podrucju Vojvodine. Acta Veterinaria, 17, 433 - 438.","Zivkovic, V. (1969) Simulide (Diptera, Simuliidae) Vojvodine. II deo. Acta Veterinaria, 19, 297 - 306.","Zivkovic, V. (1970 a) Simulide (Diptera, Simuliidae) koje napadaju coveka i domace zivotinje. Acta Veterinaria, 20, 209 - 219. Zivkovic, V. (1970 b) Trois especes de simulies (Diptera, Simuliidae) qui apparaissent en masse en Yougoslavie. Cah. O. R. S. T. O. M., ser. Ent. med. Parasitol., 8, 113 - 120.","Zivkovic, V. (1971) Black flies (Diptera, Simuliidae) of the Danube in Yugoslavia. Acta Veterinaria, 21, 225 - 236.","Sudaric Bogojevic, M., Ignjatovic Cupina, A., Petric, D. & Merdic, E. (2009) Notes on Black flies (Diptera, Simuliidae) of the Nature Park Kopacki Rit (Croatia). Proceedings of the 10 th Croatian Biological Congress with International Participation, September 14 - 20, Osijek, Croatia, pp. 211 - 212."]}

Simulium (Simulium) voilense Sherban, 1960 Literature references. River Danube at Batina (Živković 1971) (14); River Danube at Dalj (Živković 1971) (15); River Danube at Borovo (Živković 1971) (16); River Danube at Vukovar (Živković 1971) (17). Note. Analysis of polytene chromosomes of S. colombaschense revealed the existence of five cytoformes, but only one of them���corresponding to S. colombaschense described from Iron Gate of the Danube���was recorded in the Danube and its tributaries (Adler et al. 2016). One of the other cytoforms might correspond to S. voilense, but a population from the type locality of S. voilense was not studied chromosomally; therefore, the validity, identity and distribution of this species remain unclear., Published as part of Ivkovi��, Marija, K��dela, Matu�� & K��delov��, Tatiana, 2016, Blackflies (Diptera: Simuliidae) in Croatia: species richness, distribution and relationship to surrounding countries in Zootaxa 4109 (1), DOI: 10.11646/zootaxa.4109.1.2, http://zenodo.org/record/266198, {"references":["Sherban [Serban], E. N (1960) Contribution a l'etude des Simulies de la Republique Populaire Roumaine. II. Simulium voilensis n. sp. Revue de Biologie, 5, 145 - 153.","Zivkovic, V. (1971) Black flies (Diptera, Simuliidae) of the Danube in Yugoslavia. Acta Veterinaria, 21, 225 - 236.","Adler, P. H., Kudelova, T. Kudela, M, Seitz, G. & Ignjatovic-Cupina, A. (2016) Cryptic Biodiversity and the origins of pest status revealed in the macrogenome of Simulium colombaschense (Diptera: Simuliidae), history's most destructive black fly. PLOS ONE. [Published online] http: // dx. doi. org / 10.1371 / journal. pone. 0147673"]}

Simulium (Wilhelmia) lineatum (Meigen, 1804) Lierature references. River Kara��ica (Živković 1967 a, 1969) (11); River Danube at Dalj (Živković 1967 b) (15); River Danube at Borovo (Živković 1967 b) (16). New records. River Krapina (4), larvae; River Sava near the Jankomir bridge (6), larvae; River Korana near Karlovac (26), larvae; River Glina (28), larvae; River Una near Hrvatska Dubica (29), larvae; River Mirna, Istarske toplice (33), larvae., Published as part of Ivkovi��, Marija, K��dela, Matu�� & K��delov��, Tatiana, 2016, Blackflies (Diptera: Simuliidae) in Croatia: species richness, distribution and relationship to surrounding countries in Zootaxa 4109 (1), DOI: 10.11646/zootaxa.4109.1.2, http://zenodo.org/record/266198, {"references":["Meigen, J. W. (1804) Klassifikazion und beschreibung der europaischen zweiflugeligen Insekten (Diptera Linn.). Erster Band. K Reichard, Braunschweig, Germany, 314 pp. + 15 plates.","Zivkovic, V. (1967 a) Invazija Simulium erythrocephalum De Geer, 1776 (Diptera, Simuliidae) u Vojvodini. Glas Srpske Akademije Nauka i umetnosti, odeljenje medicinskih nauka, 20, 163 - 170.","Zivkovic, V. (1969) Simulide (Diptera, Simuliidae) Vojvodine. II deo. Acta Veterinaria, 19, 297 - 306.","Zivkovic, V. (1967 b) Simulide (Diptera, Simuliidae) Dunava na podrucju Vojvodine. Acta Veterinaria, 17, 433 - 438."]}

Simulium (Wilhelmia) equinum (Linneus, 1758) Literature references. River Danube at Dalj (Živković 1971) (15); tufa barrier Novakovića Brod, NP Plitvice (Ivković et al. 2012, 2014) (36); tufa barrier Kozjak-Milanovac, NP Plitvice (Ivković et al. 2012, 2014) (37). New records. River Mura at Kotoriba (1), pupae; River Korana near Karlovac (26), larvae; River Glina (28), larvae; River Mirna, Istarske toplice (33), larvae., Published as part of Ivkovi��, Marija, K��dela, Matu�� & K��delov��, Tatiana, 2016, Blackflies (Diptera: Simuliidae) in Croatia: species richness, distribution and relationship to surrounding countries in Zootaxa 4109 (1), DOI: 10.11646/zootaxa.4109.1.2, http://zenodo.org/record/266198, {"references":["Linnaeus, C. (1758) Systema naturae per regna tria naturae, secundum classes, ordines, genera, species, cum caracteribus, differentiis, synonymis, locis. 10 th ed. Stockholm, Sweden, 824 pp.","Zivkovic, V. (1971) Black flies (Diptera, Simuliidae) of the Danube in Yugoslavia. Acta Veterinaria, 21, 225 - 236.","Ivkovic, M., Kesic, M. & Stloukalova, V. (2012) Contribution to the knowledge of black fly (Diptera, Simuliidae) fauna at Plitvice lakes National Park. Natura Croatica, 21, 263 - 268.","Ivkovic, M., Kesic, M., Mihaljevic, Z. & Kudela, M. (2014) Emergence patterns and ecological preferences of hematophagous black flies along oligotrophic hydrosystem. Medical and Veterinary Entomology, 28, 94 - 102. http: // dx. doi. org / 10.1111 / mve. 12019"]}

Simulium (Simulium) reptantoides Carlsson, 1962 Literature references. River Danube at Dalj (Živković 1970 a, 1971) (15); River Danube at Vukovar (Živković 1971) (17). New record. River Sava near the Jankomir bridge (6), larvae. Note. Simulium reptantoides was in the past reported from Europe under the name S. reptans. Živković (1969) and Rivosecchi and Lipparoni (1965) recorded S. pictum as species similar to S. reptans but later accepted S. pictum as a synonym of S. reptans., Published as part of Ivkovi��, Marija, K��dela, Matu�� & K��delov��, Tatiana, 2016, Blackflies (Diptera: Simuliidae) in Croatia: species richness, distribution and relationship to surrounding countries in Zootaxa 4109 (1), DOI: 10.11646/zootaxa.4109.1.2, http://zenodo.org/record/266198, {"references":["Carlsson, G. (1962) Studies on Scandinavian black flies (Fam. Simuliidae Latr.). Opuscula Entomologica, 21 (Supplement), 1 - 280.","Zivkovic, V. (1970 a) Simulide (Diptera, Simuliidae) koje napadaju coveka i domace zivotinje. Acta Veterinaria, 20, 209 - 219. Zivkovic, V. (1970 b) Trois especes de simulies (Diptera, Simuliidae) qui apparaissent en masse en Yougoslavie. Cah. O. R. S. T. O. M., ser. Ent. med. Parasitol., 8, 113 - 120.","Zivkovic, V. (1971) Black flies (Diptera, Simuliidae) of the Danube in Yugoslavia. Acta Veterinaria, 21, 225 - 236.","Zivkovic, V. (1969) Simulide (Diptera, Simuliidae) Vojvodine. II deo. Acta Veterinaria, 19, 297 - 306.","Rivosecchi, L. & Lipparoni, L. (1965) Contributo alla conoscenza dei Simulidi Italiani XI: Alcune specie nuove per la fauna Italiana. Rivista di Parassitologia, 26, 31 - 44."]}

Simulium (Eusimulium) angustipes Edwards, 1915 Literature references. River Kara��ica (Živković 1967 a) (11); River Korana in Korana village, NP Plitvice (Ivković et al. 2012, 2014) (34); tufa barrier Novakovića Brod, NP Plitvice (Ivković et al. 2012, 2014) (36); tufa barrier Kozjak-Milanovac, NP Plitvice (Ivković et al. 2012, 2014) (37); tufa barrier Labudovac, NP Plitvice (Ivković et al. 2012, 2014) (38). New records. Spring of River Riječina (30), larvae and adult ♂; Spring of River Bolju��nica (31) larvae and adult ♂; River Suvaja near Bru��ane (44), larvae; River Korana near Karlovac (26), larvae; Skradinski buk, Krka River, NP Krka (54), larvae and adult ♂, ♀., Published as part of Ivkovi��, Marija, K��dela, Matu�� & K��delov��, Tatiana, 2016, Blackflies (Diptera: Simuliidae) in Croatia: species richness, distribution and relationship to surrounding countries in Zootaxa 4109 (1), DOI: 10.11646/zootaxa.4109.1.2, http://zenodo.org/record/266198, {"references":["Edwards, F. W. (1915) On the British species of Simulium. I. The adults. Bulletin of Entomological Research, 6, 23 - 42. http: // dx. doi. org / 10.1017 / S 0007485300043364","Zivkovic, V. (1967 a) Invazija Simulium erythrocephalum De Geer, 1776 (Diptera, Simuliidae) u Vojvodini. Glas Srpske Akademije Nauka i umetnosti, odeljenje medicinskih nauka, 20, 163 - 170.","Ivkovic, M., Kesic, M. & Stloukalova, V. (2012) Contribution to the knowledge of black fly (Diptera, Simuliidae) fauna at Plitvice lakes National Park. Natura Croatica, 21, 263 - 268.","Ivkovic, M., Kesic, M., Mihaljevic, Z. & Kudela, M. (2014) Emergence patterns and ecological preferences of hematophagous black flies along oligotrophic hydrosystem. Medical and Veterinary Entomology, 28, 94 - 102. http: // dx. doi. org / 10.1111 / mve. 12019"]}

Simulium (Simulium) reptans (Linnaeus, 1758) Literature references. Gare��nica (Mikačić 1946) (10); River Danube at Batina (Živković 1970 a) (14); River Danube at Dalj (Živković 1967 b, 1971) (15); River Danube at Borovo (Živković 1967 b, 1970 a, 1971) (16); River Danube at Vukovar (Živković 1967 b, 1971) (17); River Drava at Osijek (Živković 1969) (12); Kopački Rit (Sudarić Bogojević et al. 2009) (13). New records. River Mura at Kotoriba (1), larvae, pupae; Gojačka Dobra (23), larvae. Note. Simulium reptans was in the past recorded from Europe (except Scandinavia) also under the name S. galeratum., Published as part of Ivkovi��, Marija, K��dela, Matu�� & K��delov��, Tatiana, 2016, Blackflies (Diptera: Simuliidae) in Croatia: species richness, distribution and relationship to surrounding countries in Zootaxa 4109 (1), DOI: 10.11646/zootaxa.4109.1.2, http://zenodo.org/record/266198, {"references":["Linnaeus, C. (1758) Systema naturae per regna tria naturae, secundum classes, ordines, genera, species, cum caracteribus, differentiis, synonymis, locis. 10 th ed. Stockholm, Sweden, 824 pp.","Mikacic, D. (1946) Une invasion des Simuliides en Croatie. Veterinarski Arhiv, 16, 29 - 31.","Zivkovic, V. (1970 a) Simulide (Diptera, Simuliidae) koje napadaju coveka i domace zivotinje. Acta Veterinaria, 20, 209 - 219. Zivkovic, V. (1970 b) Trois especes de simulies (Diptera, Simuliidae) qui apparaissent en masse en Yougoslavie. Cah. O. R. S. T. O. M., ser. Ent. med. Parasitol., 8, 113 - 120.","Zivkovic, V. (1967 b) Simulide (Diptera, Simuliidae) Dunava na podrucju Vojvodine. Acta Veterinaria, 17, 433 - 438.","Zivkovic, V. (1971) Black flies (Diptera, Simuliidae) of the Danube in Yugoslavia. Acta Veterinaria, 21, 225 - 236.","Zivkovic, V. (1969) Simulide (Diptera, Simuliidae) Vojvodine. II deo. Acta Veterinaria, 19, 297 - 306.","Sudaric Bogojevic, M., Ignjatovic Cupina, A., Petric, D. & Merdic, E. (2009) Notes on Black flies (Diptera, Simuliidae) of the Nature Park Kopacki Rit (Croatia). Proceedings of the 10 th Croatian Biological Congress with International Participation, September 14 - 20, Osijek, Croatia, pp. 211 - 212."]}

Simulium (Nevermannia) angustitarse (Lundstr��m, 1911) New records. Ro��ki Slap, Krka River, NP Krka (53), larvae and adult ♂; River Vrba (56); Stream Globornica (24), larvae; upper reach of Bijela rijeka, NP Plitvice (41), larvae; River Turpinjska Rijeka (25), larvae., Published as part of Ivkovi��, Marija, K��dela, Matu�� & K��delov��, Tatiana, 2016, Blackflies (Diptera: Simuliidae) in Croatia: species richness, distribution and relationship to surrounding countries in Zootaxa 4109 (1), DOI: 10.11646/zootaxa.4109.1.2, http://zenodo.org/record/266198, {"references":["Lundstrom, C. (1911) Beitrage zur Kenntnis der Dipteren Finlands. VII. Melusinidae (Simuliidae). Acta Societatis pro Fauna et Flora Fennica, 34 (Part 12), 1 - 23 + 1 plate."]}

Simulium (Nevermannia) lundstromi (Enderlein, 1921) New record. Spring of River Bolju��nica (31), larvae., Published as part of Ivkovi��, Marija, K��dela, Matu�� & K��delov��, Tatiana, 2016, Blackflies (Diptera: Simuliidae) in Croatia: species richness, distribution and relationship to surrounding countries in Zootaxa 4109 (1), DOI: 10.11646/zootaxa.4109.1.2, http://zenodo.org/record/266198, {"references":["Enderlein, G. (1921) Neue palaarktische Simuliidaen. Sitzungsberichte der Gesellschaft naturforschender Freunde zu Berlin, 1921, 212 - 224."]}

The following format is used for the distributional data: literature references (name of the site and in parentheses, the citation of the reference and site ID); new records (name of the site and in parentheses, the site ID and the life stage in which the identifications were made, i.e., adult ♂, ♀, larvae and pupae). All the sites and their numbers are listed in Table 1.

Simulium (Simulium) variegatum Meigen, 1818 Literature references. River Korana in Korana village, NP Plitvice (Ivković et al. 2012, 2014) (34); Stream Plitvica, NP Plitvice (Ivković et al. 2012, 2014) (35); tufa barrier Novakovića Brod, NP Plitvice (Ivković et al. 2012, 2014) (36); tufa barrier Labudovac, NP Plitvice (Ivković et al. 2012, 2014) (38). New records. River Zrmanja, Pađane (46), larvae; River Butižnica (47), larvae., Published as part of Ivkovi��, Marija, K��dela, Matu�� & K��delov��, Tatiana, 2016, Blackflies (Diptera: Simuliidae) in Croatia: species richness, distribution and relationship to surrounding countries in Zootaxa 4109 (1), DOI: 10.11646/zootaxa.4109.1.2, http://zenodo.org/record/266198, {"references":["Meigen, J. W. (1818) Systematische Beschreibung der bekannten europaischen zweiflugeligen Insecten 1. Forstmann, Aachen, Germany, 332 pp.","Ivkovic, M., Kesic, M. & Stloukalova, V. (2012) Contribution to the knowledge of black fly (Diptera, Simuliidae) fauna at Plitvice lakes National Park. Natura Croatica, 21, 263 - 268.","Ivkovic, M., Kesic, M., Mihaljevic, Z. & Kudela, M. (2014) Emergence patterns and ecological preferences of hematophagous black flies along oligotrophic hydrosystem. Medical and Veterinary Entomology, 28, 94 - 102. http: // dx. doi. org / 10.1111 / mve. 12019"]}

Simulium (Nevermannia) cryophilum (Rubtsov, 1959) (complex) New record. River Bukovačka Dobra (22), larvae., Published as part of Ivkovi��, Marija, K��dela, Matu�� & K��delov��, Tatiana, 2016, Blackflies (Diptera: Simuliidae) in Croatia: species richness, distribution and relationship to surrounding countries in Zootaxa 4109 (1), DOI: 10.11646/zootaxa.4109.1.2, http://zenodo.org/record/266198, {"references":["Rubtsov, I. A. (1959 - 1964) Simuliidae (Melusinidae). In: Lindner, E. (Ed), Die Fliegen der Palaearktischen Region, 14. Simuliidae (Melusinidae), E. Schweizerbart'sche Verlagsbuchhandlung (Erwin Nagele), Stuttgart, pp. 1 - 689."]}

Murine gammaherpesvirus 68 (MHV-68) is a natural pathogen of murid rodents, which serve as hosts to Haemaphysalis concinna ticks. The occurrence of MHV-68 was investigated in a total of 47 H. concinna adult ticks collected on the vegetation in Gabcikovo, situated in south-western Slovakia (47o54´0´´N, 17o35´0´´E), from May 2013 to May 2014. DNA from ticks was purified and screened by nested PCR targeting ORF50 of MHV-68 and the copy number of virus genome in ticks was determined by a real-time PCR assay specific for ORF65. The MHV-68 incidence in questing ticks was 38.3% (18/47) and the virus genome copy number per tick varied from 2x102 to 9.6x103. In this study, MHV-68 was documented for the first time in H. concinna ticks. Results expand previous data describing the occurrence of MHV-68 in Ixodes ricinus and Dermacentor reticulatus ticks collected in Slovakia, supporting the hypothesis that MHV-68 might be a newfound pathogen in ticks.

Viviparity evolved in bacteria, plants, ˃141 vertebrate lineages (ichthyosaurs, lizards, fishes, mammals, and others), and in 11 of 44 insect orders. Live-birth cockroaches preserved with brood sac (3D recovered two times optically) included Diploptera vladimir, Diploptera savba, Diploptera gemini spp.n., D. sp.1-2, and Stegoblatta irmgardgroehni from Green River, Colorado; Quilchena, Republic; McAbee, Canada; and Baltic amber, Russia (49, 54, and 45 Ma). They evolved from rare and newly evolved Blaberidae; they radiated circumtropically, later expanded into SE Asia, and have now spread to Hawaii and the SE USA. Association of autapomorphic characters that allow for passive and active protections from parasitic insects (unique wing origami pleating identical with its egg case-attacking wasp) suggest a response to high parasitic loads. Synchronized with global reorganization of the biota, morphotype destabilization in roaches lasted approximately 11-22 Ma, including both the adaptation of novel characters and the reduction of others. Thus, while viviparity can be disadvantageous, in association with new Bauplans and/or behaviors, it can contribute to the evolution of taxa with viviparous representatives that are slightly selectively preferred.

All records of blackflies (Simuliidae) from the territory of Croatia are summarized, including previously unpublished data. The blackfly fauna of Croatia consists of 28 species. Simulium (Nevermannia) angustitarse (Lundström), Simulium (Nevermannia) cryophilum (Rubtsov) complex, Simulium (Nevermannia) lundstromi (Enderlein), Simulium (Nevermannia) vernum Macquart complex, Simulium (Simulium) argyreatum Meigen, Simulium (Simulium) bezzii (Corti) complex, Simulium paraequinum Puri and Simulium pseudequinum Séguy are reported for the first time from Croatia. Information related to the ecoregions, in which species were found and specific species traits are given. Genus Prosimulium Roubaud is represented by one species only. Genus Simulium Latreille is represented by 27 species in six subgenera, with subgenus Simulium Latreille s. str. being most species rich (13 species) and subgenera Boophthora Enderlein and Trichodagmia Enderlein represented each by only one species. Compared to the neighboring countries, the Croatian species assemblage is most similar to the fauna of Slovenia and least similar to that of Italy. The relatively low number of species, presence of several species complexes and unclear identity of other species show that further research of blackflies in Croatia is needed.

The European black fly Simulium (Simulium) colombaschense (Scopoli), once responsible for as many as 22,000 livestock deaths per year, is chromosomally mapped, permitting its evolutionary relationships and pest drivers to be inferred. The species is 12 fixed inversions removed from the standard sequence of the subgenus Simulium. Three of these fixed inversions, 38 autosomal polymorphisms, and a complex set of 12 X and 6 Y chromosomes in 29 zygotic combinations uniquely characterize S. colombaschense and reveal 5 cytoforms: 'A' in the Danube watershed, 'B' in Italy's Adige River, 'C' in the Aliakmonas River of Greece, 'D' in the Aoös drainage in Greece, and 'E' in the Belá River of Slovakia. 'C' and 'D' are reproductively isolated from one another, and 'B' is considered a cytotype of 'A,' the probable name bearer of colombaschense. The species status of 'E' cannot be determined without additional collections. Three derived polytene sequences, based on outgroup comparisons, place S. colombaschense in a clade of species composed of the S. jenningsi, S. malyschevi, and S. reptans species groups. Only cytoforms 'A' and 'B' are pests. Within the Simuliidae, pest status is reached through one of two principal pathways, both of which promote the production of large populations of blood-seeking flies: (1) colonization of the world's largest rivers (habitat specialization) or (2) colonization of multiple habitat types (habitat generalization). Evolutionary acquisition of the ability to colonize large rivers by an ancestor of the S. jenningsi-malyschevi-reptans clade set the scene for the pest status of S. colombaschense and other big-river members of the clade. In an ironic twist, the macrogenome of S. colombaschense reveals that the name associated with history's worst simuliid pest represents a complex of species, two or more of which are nonpests potentially vulnerable to loss of their limited habitat.

All records of blackflies (Simuliidae) from the territory of Croatia are summarized, including previously unpublished data. The blackfly fauna of Croatia consists of 28 species. Simulium (Nevermannia) angustitarse (Lundström), Simulium (Nevermannia) cryophilum (Rubtsov) complex, Simulium (Nevermannia) lundstromi (Enderlein), Simulium (Nevermannia) vernum Macquart complex, Simulium (Simulium) argyreatum Meigen, Simulium (Simulium) bezzii (Corti) complex, Simulium paraequinum Puri and Simulium pseudequinum Séguy are reported for the first time from Croatia. Information related to the ecoregions, in which species were found and specific species traits are given. Genus Prosimulium Roubaud is represented by one species only. Genus Simulium Latreille is represented by 27 species in six subgenera, with subgenus Simulium Latreille s. str. being most species rich (13 species) and subgenera Boophthora Enderlein and Trichodagmia Enderlein represented each by only one species. Compared to the neighboring countries, the Croatian species assemblage is most similar to the fauna of Slovenia and least similar to that of Italy. The relatively low number of species, presence of several species complexes and unclear identity of other species show that further research of blackflies in Croatia is needed.