Association between <Emphasis Type="Italic">cagA</Emphasis>, <Emphasis Type="Italic">vacAi</Emphasis>, and <Emphasis Type="Italic">dupA</Emphasis> genes of <Emphasis Type="Italic">Helicobacter pylori</Emphasis> and gastroduodenal pathologies in Chilean patients

In addition to the already known cagA gene, novel genetic markers have been associated with Helicobacter pylori (H. pylori) virulence: the dupA and vacAi genes. These genes might play an important role as specific markers to determine the clinical outcome of the disease, especially the vacAi gene, which has been expected to be a good marker of severe pathologies like gastric adenocarcinoma. In the present study, the association of cagA, dupA, and vacAi genes with gastroduodenal pathologies in Chilean patients was studied. One hundred and thirty-two patients positive for H. pylori were divided into two groups—non-severe and severe gastric pathologies—and investigated for the presence of cagA, dupA, and vacAi H. pylori virulence genes by PCR. The cagA gene was detected in 20/132 patients (15.2%), the vacAi1 gene was detected in 54/132 patients (40.9%), the vacAi2 gene was detected in 26/132 patients (19.7%), and the dupA gene was detected in 50/132 (37.9%) patients. Logistic regression model analysis showed that the vacAi1 isoform gene in the infected strains and the severity of the diseases outcome were highly associated, causing severe gastric damage that may lead to gastric cancer (p < 0.0001; OR = 8.75; 95% CI 3.54–21.64). Conversely, cagA (p = 0.3507; OR = 1.62; 95% CI 0.59–4.45) and vacAi2 (p = 0.0114; OR = 3.09; 95% CI 1.26–7.60) genes were not associated with damage, while the dupA gene was associated significantly with non-severe clinical outcome (p = 0.0032; OR = 0.25; 95% CI 0.09–0.65). In addition, dupA gene exerts protection against severe gastric pathologies induced by vacAi1 by delaying the outcome of the disease by approximately 20 years.     

Parasitism of the rhizocephalan <Emphasis Type="Italic">Briarosaccus callosus</Emphasis> on the crab <Emphasis Type="Italic">Paralomis verrilli</Emphasis> (Crustacea: Lithodidae) in the waters of southeast Sakhalin

The crab Paralomis verrilli in the waters of southeast Sakhalin is infected by the parasitic rhizocephalan barnacle Briarosaccus callosus. The prevalence of parasitic infection was on the average 4.36% (6.29% for females, 3.28% for males) and varied between samples from none up to 14.9%. The degree of prevalence was not related to the average carapace width and the sex ratio of crabs in samples. Sterilization of female P. verrilli was caused by the B. callosus infestation or its consequences. No more than two parasite externae per crab were found. Crabs with two externae made up 5.6% of all infected specimens. The infestation of crab hosts with two B. callosus externae negatively influenced the growth of the externae. A positive relationship was found between the width of the crab carapace and the length of the parasitic externae. The survival rate of P. verrilli with either one or two parasite B. callosus externae did not differ substantially. The proportion of crabs with externae and those with “scars” (12.2% in our case) can be taken as the index of survival of the parasitized crabs.     

<Emphasis Type="Italic">Drosophila melanogaster</Emphasis> as a polymicrobial infection model for <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> and <Emphasis Type="Italic">Staphylococcus aureus</Emphasis>

Non-mammalian infection models have been developed over the last two decades, which is a historic milestone to understand the molecular basis of bacterial pathogenesis. They also provide small-scale research platforms for identification of virulence factors, screening for antibacterial hits, and evaluation of antibacterial efficacy. The fruit fly, Drosophila melanogaster is one of the model hosts for a variety of bacterial pathogens, in that the innate immunity pathways and tissue physiology are highly similar to those in mammals. We here present a relatively simple protocol to assess the key aspects of the polymicrobial interaction in vivo between the human opportunistic pathogens, Pseudomonas aeruginosa and Staphylococcus aureus, which is based on the systemic infection by needle pricking at the dorsal thorax of the flies. After infection, fly survival and bacteremia over time for both P. aeruginosa and S. aureus within the infected flies can be monitored as a measure of polymicrobial virulence potential. The infection takes ~24 h including bacterial cultivation. Fly survival and bacteremia are assessed using the infected flies that are monitored up to ~60 h post-infection. These methods can be used to identify presumable as well as unexpected phenotypes during polymicrobial interaction between P. aeruginosa and S. aureus mutants, regarding bacterial pathogenesis and host immunity.     

Infestation with the parasitic nematode <Emphasis Type="Italic">Philometra ovata</Emphasis> does not impair behavioral sexual competitiveness or odor attractiveness of the male European minnow (<Emphasis Type="Italic">Phoxinus phoxinus</Emphasis>)

Our understanding on the role of chemical signals in parasite-mediated sexual selection is still limited, and only some existing studies have focused on fish. Furthermore, published studies on the effect of parasite infection on behavioral sexual competition of the male hosts have yielded contradictory results. Here, we examined whether the infection of the body cavity-dwelling parasitic nematode Philometra ovata influences odor-based female choice and behavioral sexual competition (dominance and courtship behavior) between males in the European minnow (Phoxinus phoxinus), the cyprinid fish host. In contrast to our predictions, we found that naïve females showed no preference between the odors of infected and non-infected males, thus indicating that P. ovata infection may not affect odor-based female choice. Moreover, P. ovata did not impair sexual competitiveness of their hosts either. Our results indicate that despite its relatively large size, P. ovata may not alter sexual cues and the success of the male hosts in sexual selection.     

Preliminary data on the variability of three microsatellite loci in Pacific <Emphasis Type="Italic">Gadus macrocephalus</Emphasis> and Atlantic <Emphasis Type="Italic">G. morhua</Emphasis> cod (Gadidae)

Variability of microsatellite DNA loci Gmo3, Gmo34, and Gmo35 is studied in samples of Pacific cod Gadus macrocephalus and Atlantic cod G. morhua. The results show high values of identity of the samples within the North Pacific basin (0.9766–0.9924) and within the Northeast Atlantic basin (0.9580). Based on the pairwise assessment of genetic differentiation, the F _ST values are significantly different in all variants between the samples of Pacific and Atlantic cod (F _ST = 0.5235–0.6719, p < 0.001). Within the basins, the significant differences in the frequencies of main alleles are revealed in the loci Gmo3 and Gmo34 for the samples from the Pacific and Atlantic oceans, respectively.     

qPCR estimates of <Emphasis Type="Italic">Babesia bovis</Emphasis> and <Emphasis Type="Italic">Babesia bigemina</Emphasis> infection levels in beef cattle and <Emphasis Type="Italic">Rhipicephalus microplus</Emphasis> larvae

Babesia spp. are tick-transmitted intraerythrocytic apicomplexan parasites that infect wild and domestic animals. Babesia bovis and B. bigemina are endemic and responsible for enormous economic losses to the livestock industry in most of the Brazilian territory, wherein the tick Rhipicephalus microplus is the unique vector. Better understanding of epidemiology and parasite–host interactions may improve the tools for disease control and genetic management for selection of resistant animals. This study aimed to detect, quantify and measure the correlation between B. bigemina and B. bovis infection levels in bovine blood and into tick, by absolute quantification of hemoparasite DNA using qPCR. Blood bovine samples and larvae pools from 10 engorged R. microplus females were collected from each Canchim heifers (5/8 Charolais?+?3/8 zebu, n?=?36). All evaluated samples were positive for both Babesia species tested. Correlations of B. bovis and B. bigemina levels between cattle and tick host were 0.58 and 0.66, respectively. These high positive correlation coefficients indicate that parasitemia load in the bovine may be dependent on or may determine the parasitemia load in the ticks.     

Usefulness of the Non-conventional <Emphasis Type="Italic">Caenorhabditis elegans</Emphasis> Model to Assess <Emphasis Type="Italic">Candida</Emphasis> Virulence

Invasive candidiasis is caused mainly by Candida albicans, but other Candida species have increasing etiologies. These species show different virulence and susceptibility levels to antifungal drugs. The aims of this study were to evaluate the usefulness of the non-conventional model Caenorhabditis elegans to assess the in vivo virulence of seven different Candida species and to compare the virulence in vivo with the in vitro production of proteinases and phospholipases, hemolytic activity and biofilm development capacity. One culture collection strain of each of seven Candida species (C. albicans, Candida dubliniensis, Candida glabrata, Candida krusei, Candida metapsilosis, Candida orthopsilosis and Candida parapsilosis) was studied. A double mutant C. elegans AU37 strain (glp-4;sek-1) was infected with Candida by ingestion, and the analysis of nematode survival was performed in liquid medium every 24 h until 120 h. Candida establishes a persistent lethal infection in the C. elegans intestinal tract. C. albicans and C. krusei were the most pathogenic species, whereas C. dubliniensis infection showed the lowest mortality. C. albicans was the only species with phospholipase activity, was the greatest producer of aspartyl proteinase and had a higher hemolytic activity. C. albicans and C. krusei caused higher mortality than the rest of the Candida species studied in the C. elegans model of candidiasis.     

Feeding of Polar cod <Emphasis Type="Italic">Boreogadus saida</Emphasis> in the Kara Sea

In the summer–autumn period of 2007, 2010, and 2013, the peculiarities of Polar cod feeding Boreogadus saida in the Kara Sea were considered. In these years, the intensity of feeding of adult individuals was quite similar. Spatial and interannual differences in the composition of the food have been observed. Copepoda, Euphausiacea, and tunicates (Oikopleura sp.) dominated in the food of young fish. The main preys of adult individuals are Hyperiidea, Copepoda, Euphausiacea, and fish. As the length of Polar cod increases, the weight percentage of copepods decreases and the portion of fish increases.     

Molecular characterization and expression analysis of <Emphasis Type="Italic">Hsp90</Emphasis> in <Emphasis Type="Italic">Schizothorax prenanti</Emphasis>

Aquatic animals suffer from various environmental stresses because the aquatic environment is a very complex system. To monitor the health status of fish, Hsp90 a potential early warning marker was determined in Schizothorax prenanti after infection with a bacterium. In this study, we cloned Hsp90 from S. prenanti for the first time. The full-length cDNA sequence of SpHsp90 was 2663 bp, contains an open reading frame of 2181 bp, and has a gene encoding 726 amino acids, an estimated molecular mass of 83.38 kDa, and a theoretical isoelectric point of 4.91. The SpHsp90 amino acid sequence has five conserved HSP90 family signatures and shares 87.0–95.5 % identity with other vertebrates. Phylogenetic analysis and structure comparison indicated that SpHsp90 should be a β isoform of the HSP90 family. SpHsp90 was ubiquitously expressed in all examined tissues, and the highest level of expression was in the kidney. After Streptococcus agalactiae infection, the level of SpHsp90 expression had significant changes (P < 0.05) in the hepatopancreas, spleen, kidney, and blood. The expression increased to the highest level at 6 h in the blood and at 24 h in the hepatopancreas, spleen, and kidney. The results suggested that the SpHsp90 gene could be induced by S. agalactiae in S. prenanti and that SpHsp90 may be involved in resistance to bacterial infection and provide an early warning information. The kidney is the most suitable for detecting SpHsp90 after bacterial infection.     

Occurrence of the tuna nematode <Emphasis Type="Italic">Hysterothylacium cornutum</Emphasis> (Stossich, 1904) in farmed Atlantic cod <Emphasis Type="Italic">Gadus morhua</Emphasis> L. in North Norway

Adult and fourth-stage larval nematodes found in the stomachs of farmed cod in North Norway in October 2006 were identified as Hysterothylacium cornutum (Stossich, 1904), a nematode considered to be specific at the adult stage to tunas of the genus Thunnus. As far as we are aware, this is the first report of an adult form of this nematode from any host other than members of the genus Thunnus, and is also the first report from a polar region. The two infected cod, one with one large adult worm and another with six larvae, were in a sample of 17 that had been captured from the wild about 1 year before sampling and held in floating sea cages in Øksfjord, Finnmark County, North Norway, where the examinations took place. No further infections were found in any other samples of wild and farmed cod, totalling 261 fish, examined by us during the period 2006 and 2007 from five locations distributed along the coast of Norway from Øksfjord in the north to Ålesund in the south. Possible sources of this unusual infection are discussed, but no firm conclusion could be reached.     

Association between expression levels and growth trait-related SNPs located in promoters of the <Emphasis Type="Italic">MC4R</Emphasis> and <Emphasis Type="Italic">MSTN</Emphasis> genes in <Emphasis Type="Italic">Spinibarbus hollandi</Emphasis>

Melanocortin 4 receptor: (MC4R) and Myostatin (MSTN) are two important growth trait-related genes in animals. In this study, we showed that two SNPs, MC4R-719A>G and MSTN-519C>T, found in the promoters of the MC4R and MSTN genes, respectively, are both associated with growth traits in Spinibarbus hollandi. Furthermore, we observed that there were significant associations between the expression levels of the MC4R and MSTN genes and these two growth trait-related SNPs. The expression level of MC4R gene in brain was lower in GG genotype fish with extremely high growth performance than that in AA genotype fish with extremely low growth performance. Expression level of the MSTN gene in muscle was lower in TT genotype fish with extremely high growth performance than that in CC and CT genotype fish with lower growth performance. The results indicated that these SNPs located in the promoters of MC4R and MSTN are associated with growth-related traits through modification of gene expression levels. The MSTN and MC4R SNPs may have useful application in effective marker-assisted selection aimed to increase output in S. hollandi.     

Immunohistochemical Cross-Reactivity Between <Emphasis Type="Italic">Paracoccidioides</Emphasis> sp. from Dolphins and <Emphasis Type="Italic">Histoplasma capsulatum</Emphasis>

Paracoccidioidomycosis ceti is a cutaneous disease of cetaceans caused by uncultivated Paracoccidioides brasiliensis or Paracoccidioides spp. Serological cross-reactions between paracoccidioidomycosis ceti and paracoccidioidomycosis, paracoccidioidomycosis and histoplasmosis, and paracoccidioidomycosis and coccidioidomycosis have been reported before. The present study aimed to detect immunohistochemical cross-reaction between antibodies to Paracoccidioides sp. and Histoplasma capsulatum, and vice versa. Thirty murine sera, obtained from experimental infections of 6 isolates of H. capsulatum, were reacted with paraffin-embedded yeast-form cells of Paracoccidioides sp. derived from a case of paracoccidioidomycosis ceti in Japan. The murine sera were also reacted with human isolates of H. capsulatum yeast cells, with P. brasiliensis yeast cells, and with fungal cells of Coccidioides posadasii. Three dolphins’ sera from cases of paracoccidioidomycosis ceti, two human sera from patients with paracoccidioidomycosis, and a serum from a healthy person with a history of coccidioidomycosis were used in order to determine that the tested fungal cells reacted properly. Sera derived from mice infected with an isolate of H. capsulatum reacted positively against yeast cells of Paracoccidioides sp., yeast cells of P. brasiliensis, and fungal cells of C. posadasii, while those derived from other strains were negative. The present study recorded for the first time the cross-reaction between the yeast cells of H. capsulatum and antibodies against Paracoccidioides spp., the yeast cells of Paracoccidioides sp. and antibodies against H. capsulatum, the yeast cells of Paracoccidioides sp. and antibodies against Coccidioides sp., and fungal cells of C. posadasii and antibodies against Paracoccidioides spp.     

Molecular phylogeny of pectinase genes <Emphasis Type="Italic">PGU</Emphasis> in the yeast genus <Emphasis Type="Italic">Saccharomyces</Emphasis>

Using yeast genome databases and literature data, phylogenetic analysis of pectinase PGU genes from 112 Saccharomyces strains assigned to the biological species S. arboricola, S. bayanus (var. uvarum), S. cariocanus, S. cerevisiae, S. kudriavzevii, S. mikatae, S. paradoxus, and the hybrid taxon S. pastorianus (syn. S. carlsbergensis) was carried out. A superfamily of divergent PGU genes was found. Natural interspecies transfer of the PGU gene both from S. cerevisiae to S. bayanus and from S. paradoxus to S. cerevisiae may, however, occur. Within the Saccharomyces species, identity of the PGU nucleotide sequences was 98.8–100% for S. cerevisiae, 86.1–95.7% for S. bayanus (var. uvarum), 94–98.3% for S. kudriavzevii, and 96.8–100% for S. paradoxus/S. cariocanus. For the first time, a family of polymeric PGU1b, PGU2b, PGU3b and PGU4b genes is documented for the yeast S. bayanus var. uvarum, a variety important for winemaking.     

<Emphasis Type="Italic">Myxobolus imparfinis</Emphasis> n. sp. (Myxozoa: Myxosporea), a new gill parasite of <Emphasis Type="Italic">Imparfinis mirini</Emphasis> Haseman (Siluriformes: Heptapteridae) in Brazil

A new species of myxozoan, Myxobolus imparfinis n. sp. is described based on material from the gills of Imparfinis mirini (Haseman) (Heptapteridae). Mature myxospores are round, measuring 7.1–8.4 (7.9 ± 0.3) μm in length, 4.5–6.2 (5.5 ± 0.5) μm in width and 3.1–4.2 (3.7 ± 0.3) μm in thickness. The polar capsules are of unequal size, the larger polar capsule measuring 3.4–4.5 (3.9 ± 0.3) μm in length and 1.4–2.0 (1.7 ± 0.1) μm in width and the smaller capsule measuring 3.1–3.8 (3.4 ± 0.2) μm in length and 1.2–1.8 (1.5 ± 0.2) μm in width. The polar filament presents 6–7 coils. Spores had a prevalence of infection of 75% (6/8). In histological analyses we detected the development site of spores in primary filaments, in afferent branchial artery, thus classifying the type of infection to the filamental type and vascular subtype. The phylogenetic analyses of a dataset including species Myxobolus Bütschli, 1882 and Henneguya Thélohan, 1892 from South America recovered M. imparfinis n. sp. as a sister species of Myxobolus flavus Carriero, Adriano, Silva, Ceccarelli & Maia, 2013. To our knowledge, this is the first record of a myxozoan species parasitising I. mirini.     

A delayed effect of the aquatic parasite <Emphasis Type="Italic">Margaritifera laevis</Emphasis> on the growth of the salmonid host fish <Emphasis Type="Italic">Oncorhynchus masou masou</Emphasis>

Parasitic species often have detrimental effects on host growth and survival. The larvae of the genus Margaritifera (Bivalvia), called glochidia, are specialist parasites of salmonid fishes. Previous studies have reported negligible influences of the parasite on their salmonid hosts at natural infection levels. However, those studies focused mainly on their instantaneous effects (i.e., during the parasitic period). Given the time lag between physiological and somatic responses to pathogen infections, the effect of glochidial infection may become clearer during the post-parasitic period. Here, we examined whether the effect of glochidial infections of Margaritifera laevis on its salmonid host Oncorhynchus masou masou would emerge during the post-parasitic period. We performed a controlled aquarium experiment and monitored fish growth at two time intervals (i.e., parasitic and post-parasitic periods) to test this hypothesis. Consistent with previous observations, the effects of glochidial infection were unclear in the middle of the experiment (day 50; parasitic period). However, even with a natural glochidial load (48 glochidia per fish), we found a significant reduction in growth rates of infected fish in the extended period of the experiment (day 70; post-parasitic period). Our results suggest that examining only instantaneous effects may provide misleading conclusions about mussel–host relationships.     

Occurrence of a gelatinous predator (<Emphasis Type="Italic">Cyanea capillata</Emphasis>) may affect the distribution of <Emphasis Type="Italic">Boreogadus saida</Emphasis>, a key Arctic prey fish species

Although the Arctic cod (Boreogadus saida) has a pan-Arctic distribution, little is known about its occurrence in near-shore waters where this species is the principal prey for seabirds, marine mammals and other fish. Published research describes the scyphomedusa Cyanea capillata as an Arctic cod predator, and this paper presents observations from long-term investigations using active hydroacoustics that suggest the Arctic cod avoided C. capillata in two small bays of Cornwallis Island (Canadian High Arctic archipelago). Distribution patterns in echograms suggested that features such as boundary layer fronts restricted jellyfish movements and Arctic cod were often abundant on the side of fronts where C. capillata were absent. Thus, habitat partitioning allowed Arctic cod to share habitat with its predator, albeit exceptions to this sharing occurred when jellyfish abundance was high and Arctic cod were displaced. Thus, if a warmer Arctic triggers an increase in C. capillata abundance, it is possible that small-scale aspects of Arctic cod distribution could be affected. This in turn could have significant ripple effects within the Arctic food web, an additional and previously unrecognized consequence of climate change.     

Revision of the genus <Emphasis Type="Italic">Melamphaes</Emphasis> (Melamphaidae). 5. Oligo-raker species: <Emphasis Type="Italic">M. indicus</Emphasis>, <Emphasis Type="Italic">M. eurous</Emphasis>, and <Emphasis Type="Italic">M. typhlops</Emphasis>

Three oligo-raker species (?19 rakers on the first gill arch) of the genus Melamphaes out of the “M. typhlops” group are considered. The validity of M. indicus Ebeling is restored. This species inhabits equatorial and tropical waters of the Indian Ocean and the western part of the Pacific Ocean. M. eurous sp. n., which is related to M. indicus, is described from equatorial waters of the eastern part of the Pacific Ocean. M. typhlops (Lowe) inhabiting the northern part of the Atlantic Ocean, from the equatorial zone about to 45° N, is redescribed.