Some ecological aspects of the copepod parasites of the common hake, Merluccius hubbsi, from the Argentine-Uruguayan coasts

A total of 2164 common hake Merluccius hubbsi captured in the Argentine-Uruguayan Common Fishing Zone was examined for parasitic copepods. The fish were infested with Chondracanlhus palpifer and Neobrachietia insidiosa f. lageniformis, the former parasitizing the buccal cavity and the latter the branchial arches. C. palpifer showed an increase in both prevalence and intensity in relation to the host size; infestation with N. insidiosa f. lageniformis decreased with increasing length of hake. Attachment site preferences and variations in the parasite distribution patterns as related to the host biology were observed. Evidence of negative association between copepod species and seasonal changes in the parasite composition was also found.     

Ingestion of the dinoflagellate, Pfiesteria piscicida, by the calanoid copepod, Acartia tonsa

The dinoflagellate, Pfiesteria piscicida, can form harmful algal blooms in estuarine environments. The dominant copepod species usually found in these waters is Acartia tonsa. We tested the ability of A. tonsa to graze the non-toxic zoospore stage of P. piscicida and thus serve as a potential biological control of blooms of this algal species. A. tonsa grazed the non-toxic zoospore stages of both a non-inducible P. piscicida strain (FDEPMDR23) and a potentially toxic strain (Tox-B101156) at approximately equal rates. Ingestion of P. piscicida increased with cell concentration and exhibited a saturated feeding response. Both the maximum number of cells ingested (I_max) and the slope of the ingestion curve (α) of A. tonsa feeding on P. piscicida were comparable to these ingestion parameters for A. tonsa fed similar-sized phytoplankton and protozoan species. When these laboratory ingestion rates were combined with abundance estimates of A. tonsa from the Pocomoke Estuary and Chesapeake Bay, we found that significant grazing control of the non-toxic zoospore stage of P. piscicida by A. tonsa would only occur at high copepod abundances (>10 copepods L⁻¹). We conclude that under most in situ conditions the potential biological control of blooms of P. piscicida is exerted by microzooplankton grazers. However, in the less saline portions of estuaries where maximum concentrations of copepods often occur with low abundances of microzooplankton, copepod grazing coefficients can be similar to the growth rates of P. piscicida.     

Prorocentrum minimum(clone Exuv) is nutritionally insufficient, but not toxic to the copepod Acartia tonsa

This study tested whether the dinoflagellate Prorocentrum minimum is nutritionally insufficient or toxic to the copepod Acartia tonsa. Experiments were carried out with adult female A. tonsa and the P. minimum clone Exuv, both isolated from Long Island Sound. Initially, the functional and numerical responses of A. tonsa feeding on exponentially growing P. minimum cells were characterized. These experiments revealed that A. tonsa readily ingested P. minimum cells, up to the equivalent of 200% of body carbon day⁻¹, but egg production was relatively low, with a maximum egg production rate of 22% of body carbon day⁻¹. Hence, the egg production efficiency (egg carbon produced versus cell carbon ingested) was low (10%). In a separate experiment, ingestion and egg production rates were measured as a function of food concentration with cells in different growth stages (early-exponential, late-exponential/early-stationary, and late-stationary growth phase) to simulate conditions during a bloom. There was no indication that cells in the stationary phase resulted in lower ingestion or egg production rates relative to actively growing cells. Egg hatching success remained high (>80%) and independent of the cell growth phase. In a third experiment specifically designed to test the hypothesis that P. minimum is toxic, ingestion, egg production and egg hatching success were measured when females were fed mixtures of P. minimum and the diatom Thalassiosira weissflogii, but in which total food concentration was held constant and the proportion of P. minimum in the mixed diet varied. A. tonsa readily ingested P. minimum when it was offered in the mixed diet, with no detrimental effects on egg production or egg hatching observed. Supplementing P. minimum with T. weissflogii increased both the egg production rate and the egg production efficiency. It is concluded that P. minimum is nutritionally insufficient, but not toxic to A. tonsa. Finally, it is estimated that in the field grazing by A. tonsa is approximately equivalent to 30% of the maximum daily growth rate of P. minimum. Hence, copepod grazing cannot be ignored in field and modeling studies of the population dynamics of P. minimum.     

Differential responses of populations of the copepod Acartia hudsonica to toxic and nutritionally insufficient food algae

Nutritional insufficiency and toxicity are deleterious effects of phytoplankton on grazers. We hypothesize that toxic food is likely to have stronger evolutionary selective effects on grazers than nutritionally insufficient food. We explore this hypothesis in comparative studies of egg production and egg hatching of the copepod Acartia hudsonica challenged with both a toxic and a nutritionally insufficient alga. Experiments lasting 6 days, in which mixtures of different proportions of the suspect and a control alga were offered as food to female copepods, showed that the dinoflagellate Alexandrium fundyense, which bears paralytic shellfish toxins, was toxic to A. hudsonica. In contrast, the diatom Phaeodactylum tricornutum was nutritionally insufficient to A. hudsonica. In another set of experiments, the effects of A. fundyense and P. tricornutum, respectively, as sole foods on egg production and egg hatching success of two geographically separated populations (Maine and Connecticut) of the copepod A. hudsonica were examined in common-environment experiments, after being raised under identical conditions for two generations. The location in Maine regularly experiences toxic blooms of Alexandrium sp. whereas the location in Connecticut does not. During a 6-day period, A. fundyense reduced the egg production rates of the Connecticut copepod population, but not of the Maine population. In contrast, the diatom P. tricornutum reduced the egg production of both populations. These results of this study are consistent with the hypothesis of local adaptation to toxic food, but not to nutritionally insufficient food.     

Changes in lipids and biochemical properties of actomyosin from pre- and post-spawned hake (Merluccius hubbsi Marini)

The total lipid contents of muscle, the liver somatic index and the total lipid contents of actomyosin in hakes changed with the gonadal condition of the fish. Non-polar lipids in actomyosin from pre-spawned hake were 145% higher than in actomyosin from post-spawned hake; polar lipids were 30% higher. The relative percentage of phospholipids changed from 32% in pre-spawned hake to 48% in post-spawned hake. The Mg²⁺-ATPase activity in actomyosin increased from the pre-spawned to the post-spawned conditions, and this could be related to a higher phospholipids to neutral lipids ratio in post-spawned hake. Gradual decreases in both Ca²⁺-ATPase activity and myosin: actin ratio during the gonadal development of fish were found, suggesting a partial loss of myosin functionality of the actomyosin complex.     

New species of Phaeoramularia, Pseudocercospora and Stenella from Western Ghats of India

Four new species of the hyphomycete genera Phaeoramularia viz. Ph. caesalpinae, Pseudocercospora viz., Ps. tiliacearum, Stenella, viz. S. argyreiae and S. grewiae occurring on Caesalpinia bonducella Fleming (Caesalpiniaceae), Grewia sp. (Tiliaceae), Argyrea sp. Lour (Convolvulaceae) and Grewia sp. L. (Tiliaceae), respectively are described and illustrated here. All these fungi were collected from Western Ghats of India.     

Host range of an NPV and a GV isolated from the common cutworm, Agrotis segetum: pathogenicity within the cutworm complex

The term cutworm covers a range of species with a similar life history that can be very damaging pests on a wide range of crops. Attacks by cutworms are often made up of more than one species; thus, the most cost effective microbial control agent needs to be pathogenic for multiple species within this complex. In this study we investigate the host range of Agrotis segetum NPV and A. segetum GV for other cutworm species and closely related Noctuinae. Eight species, A. segetum, Agrotis ipsilon, Agrotis exclamationis, Agrotis puta, Noctua comes, Peridroma saucia, Xestia sexstrigata, and Xestia xanthographa, were clearly susceptible to AgseNPV, which was confirmed by DNA analysis. Aglais urticae, Diarsia rubi, Noctua pronuba, and Xestia c-nigrum were not susceptible to AgseNPV at the doses used. Noctua fimbriata, Noctua janthina, and Ochroplura plecta gave ambivalent results: larvae died of NPV infection when they were challenged with AgseNPV, but these individuals only produced weak positives in a squash blot analysis and there was insufficient DNA for confirmation by restriction endonuclease profiling. These ambivalent results could suggest either a weak infection by AgseNPV or partial homology between their own virus and AgseNPV. The untreated control insects of several species died of NPV infection, which indicates that these field-collected insects were probably carrying a vertically transmitted NPV. Fewer species were tested with AgseGV and only N. pronuba and N. comes were susceptible. N. fimbriata and Helicoverpa armigera were not susceptible to AgseGV.     

Main sterols from the ophiuroids Ophiocoma echinata, Ophiocoma wendtii, Ophioplocus januarii and Ophionotus victoriae

Sterol compositions of the cold water ophiuroids Ophioplocus januarii and Ophionotus victoriae and of the tropical ophiuroids Ophiocoma echinata and Ophiocoma wendtii are reported. The four sterol mixtures contain Δ⁵ mono- and di-unsaturated common 3β-hydroxy-sterols. Ophioplocus januarii and O. victoriae contain 24-methylcholesta-5,24(28)-dien-3β-ol and 24ζ-ethylcholesta-5,24(28)-dien-3β-ol in higher abundance than in O. echinata. These sterols were not found in O. wendtii. An interesting finding is the presence of Δ^5,24(28)-24-n-propylidenecholesterol in 7.6% in Ophionotus victoriae.     

The phylogenetic placement of the Leucogastrales, including Mycolevis siccigleba (Cribbeaceae), in the Albatrellaceae using morphological and molecular data

The morphology of many hypogeous fungi converges on a homogeneous reduced form, suggesting that disparate lineages are subject to a uniform selection pressure. The primary goal of this study was to evaluate the morphology and infer the phylogeny of the Leucogastrales with Mycolevis siccigleba using a Bayesian methodology. A comprehensive morphological assessment was used for an a priori phylogenetic inference to guide the sequencing effort. All structures except spore ornamentation pointed to the Albatrellaceae as the most likely sister taxon. Polyporoletus sublividus, a close relative of Albatrellus, produces ornamented basidiospores with a similar structure to M. siccigleba basidiospores. The ITS from 30 taxa was used for the molecular phylogenetic analysis. P. sublividus was found sister to Mycolevis. Leucophleps spinispora and L. magnata formed a group sister to the Polyporoletus/Mycolevis group, whereas Leucogaster was polyphyletic with respect to the core of the Leucogastrales and sister to A. caeruleoporus. This relationship was expected as previously undescribed chlamydospores produced by members of Albatrellus had a similar morphology to the basidiospores of L. rubescens.     

Evolutionary relationships among aquatic anamorphs and teleomorphs: Lemonniera, Margaritispora, and Goniopila

The hypothesis that similar conidial morphologies in aquatic hyphomycetes are a result of convergent evolution was tested using molecular sequence data. Cladistic analyses were performed on partial sequences of 28S rDNA of seven species of Lemonniera, one species of Margaritispora and one species of Goniopila. Lemonniera has tetraradiate conidia with long arms, whereas Margaritispora and Goniopila have typically globose (isodiametric) conidia, with short conical protuberances in a stellate or quadrangular arrangement. Lemonniera and Margaritispora have phialidic conidiogenesis and both produce dark, minute sclerotia in culture whereas Goniopila has holoblastic conidiogenesis and does not produce sclerotia in culture. Goniopila produces a microconidial phialidic synanamorph in culture. All three genera have schizolytic conidial secession. Molecular analyses demonstrate that Lemonniera species are placed in two distinct clades: one within Leotiomycetes; the other within Pleosporales, Dothideomycetes. Margaritispora is placed with Lemonniera species within Leotiomycetes. Goniopila and Lemonniera pseudofloscula are placed within Dothideomycetes. No morphological character was entirely congruent with the molecular derived phylogeny. This suggests that for the group of species studied, conidial shape is not a reliable indicator of phylogeny but more likely the result of convergent evolution in response to the aquatic environment.     

Parasitoids associated with the common pistachio psylla, Agonoscena pistaciae, in Iran

The parasitoids associated with the common pistachio psylla, Agonoscena pistaciae Burckhardt and Lauterer, were investigated at three pistachio plantations in Rafsanjan, Iran. Of the 6504 wasps emerging from mummified psyllids, 46% were the primary parasitoid Psyllaephagus pistaciae Ferrière, and the remaining 54% represented six species of hymenopterous hyperparasitoids, including Chartocerus kurdjumovi (Nikol’skaja), Marietta picta (André), Pachyneuron aphidis (Bouché), Pachyneuron muscarum (Linnaeus), Psyllaphycus diaphorinae (Hayat), and Syrphophagus aphidivorus (Mayr). Lysiphlebus fabarum Marshall, the parasitoid of Aphis gossypii Glover and Aphis craccivora Koch present on weeds, was found to be an alternative host for three major hyperparasitoids of A. pistaciae. The most abundant hyperparasitoid was S. aphidivorus, appearing during the growing season in all trial locations on psyllids and aphids in pistachio orchards. The weed-infesting aphids, along with their primary parasitoid, can act as a reservoir of A. pistaciae secondary parasitoids. Therefore, parasitized aphids allow populations of secondary parasitoids to increase and consequently to apply higher pressure on P. pistaciae. We detected that two primary parasitoid species, including P. pistaciae and L. fabarum, attacking different species of hosts interact indirectly through shared secondary parasitism. It is suggested that the community structure of A. pistaciae may be influenced by apparent competition, although more work is needed to provide firm evidence.     

A new fungal genus, Teracosphaeria, with a phialophora-like anamorph (Sordariomycetes, Ascomycota)

The new genus and species Teracosphaeria petroica is described for a perithecial ascomycete and its anamorph occurring on decayed wood collected in New Zealand. The fungus produces immersed, non-stromatic ceratosphaeria-like perithecia in nature, with hyaline, septate ascospores produced in unitunicate, non-amyloid asci. The anamorph produced in vitro is phialophora-like with lightly pigmented phialides terminating in flaring, deep collarettes that are often noticeably brown with conspicuous periclinal thickening. Phylogenetic analysis of LSU rDNA sequence data indicates that this fungus is distinct from morphologically similar fungi classified in the Chaetosphaeriales, the Trichosphaeriales or the Magnaporthaceae. It forms a monophyletic group with recently described, chaetosphaeria-like ascomycetes, such as the pyrenomycete genus Mirannulata, and shows affinity with the anamorphic species Dictyochaeta cylindrospora. The usefulness of describing anamorph genera for morphologically reduced anamorphs, when anamorph characteristics are actually part of the holomorph diagnosis, is discussed. An apparently contradictory example of the so-called Cordana and Pseudobotrytis anamorphs of Porosphaerella spp. is also discussed.     

三孢布拉霉crgA启动子的克隆和活性分析

【背景】CrgA是三孢布拉霉(Blakesleatrispora,Bt)中调控类胡萝卜素合成的关键负调控因子,其表达水平会影响类胡萝卜素的合成。【目的】克隆三孢布拉霉crgA启动子并分析其活性,为进一步解析CrgA表达调控机制奠定基础。【方法】通过综合微生物基因组(integrated microbial genomes, IMG)数据库提供的基因组序列,克隆crgA翻译起始位点上游2 000 bp序列,分析其顺式调控元件和转录起始区域预测,通过RT-qPCR分析不同光照时间对三孢布拉霉crgA相对转录水平的影响;构建4个不同长度的crgA启动子截短序列驱动的GUS-mGFP5重组表达载体p1303-procrgAF、F1、F2和F3,利用农杆菌侵染整合到三孢布拉霉基因组中,在黑暗和光照条件下测定β-D-葡萄糖苷酸酶(β-D-glucuronidase,GUS)酶活性并观察荧光信号。【结果】crgA启动子不仅包含基础的TATA-box和CAAT-box元件,还包括多个与光响应相关的元件。观察荧光结果显示CaMV35S和构建的4个突变启动子均能在三孢布拉霉体内驱动下游基因表达,检测GUS...     

马齿苋内生菌橘青霉和波兰青霉中抗青枯菌的活性物质

为了挖掘农作物病害生物防治新资源,以药用植物马齿苋(Portulaca oleracea)为材料,通过培养基种植法分离和纯化其根、茎、叶中的内生菌,以青枯菌(Ralstonia solanacearum)的抑菌试验评价其活性,采用菌落形态观察和ITS序列分析鉴定菌种。结果表明,从马齿苋筛选出2种具有抑制青枯菌的内生菌橘青霉(Penicillium citrinum)和波兰青霉(P. polonicum),采用液相与四极杆飞行时间串联质谱(UPLC-QTOF-MS)鉴定2种内生菌的主要活性物质为橘霉素,其对青枯菌的抑制效果比链霉素更好。因此,这为植物青枯病的生物防治提供科学依据。     

Energetic stress in the honeybee Apis mellifera from Nosema ceranae infection

Parasites are dependent on their hosts for energy to reproduce and can exert a significant nutritional stress on them. Energetic demand placed on the host is especially high in cases where the parasite-host complex is less co-evolved. The higher virulence of the newly discovered honeybee pathogen, Nosema ceranae, which causes a higher mortality in its new host Apis mellifera, might be based on a similar mechanism. Using Proboscis Extension Response and feeding experiments, we show that bees infected with N. ceranae have a higher hunger level that leads to a lower survival. Significantly, we also demonstrate that the survival of infected bees fed ad libitum is not different from that of uninfected bees. These results demonstrate that energetic stress is the probable cause of the shortened life span observed in infected bees. We argue that energetic stress can lead to the precocious and risky foraging observed in Nosema infected bees and discuss its relevance to colony collapse syndrome. The significance of energetic stress as a general mechanism by which infectious diseases influence host behavior and physiology is discussed.     

Candidatus Monilibacter spp., common bulking filaments in activated sludge, are members of Cluster III Defluviicoccus

Two alphaproteobacterial Neisser negative ‘Nostocoida limicola’ morphotypes differing slightly in their trichome diameter and filament regularity were dominant populations in the Bendigo, Victoria, Australia activated sludge community removing phosphorus (P). Neither responded to the FISH probes available for any of the other alphaproteobacterial ‘N. limicola’ morphotypes. Instead both fluoresced with the DF988 FISH probe designed originally to target alphaproteobacterial cluster II Defluviicoccus tetrad forming organisms. A 16S rRNA based clone library from this biomass revealed that the alphaproteobacterial clones grouped closely with Candidatus ‘Monilibacter batavus’ and Defluviicoccus clones in a cluster separate from the existing cluster I and II Defluviicoccus. When a FISH probe was designed against these, it only hybridized to the thinner and less abundant ‘N. limicola’ morphotype. Micromanipulation–RT-PCR was used to selectively recover the main ‘N. limicola’ morphotype and a FISH probe designed against the 16S rRNA clones generated from it showed only this filament fluoresced. From FISH based surveys, both ‘N. limicola’ variants occurred frequently in phosphorus removal activated sludge systems in Australia treating domestic waste. The data suggest that they represent two new strains of Candidatus ‘Monilibacter’, which on this evidence are filamentous members of the genus Defluviicoccus, a potential competitor for the polyphosphate accumulating organisms in these communities.     

Cloning and expression of ecdysone receptor (EcR) from the intertidal copepod, Tigriopus japonicus

Ecdysteroids are steroid hormones that play an important role in development, growth, molting of larva, and reproduction in the Arthropoda. The effect of ecdysteroids is mediated by its binding to ecdysteroid receptor (EcR). To investigate the role of EcR during development and the effect to environmental stressors on EcR expression in a copepod, we isolated and characterized cDNA and 5′-promoter region of the Tigriopus japonicus EcR (TJ-EcR), and studied mRNA expression pattern. The full-length TJ-EcR cDNA sequence was 1962 bp in length and the open reading frame encoded 546 amino acids. The deduced TJ-EcR protein contained well-conserved DNA-binding domain and ligand-binding domain. Phylogenetic analysis revealed that TJ-EcR was clustered with the EcR of other crustaceans. TJ-EcR mRNA was expressed in a developmental stage-specific manner: high in early developmental stages and low in the adult stage. Significantly elevated expression of the TJ-EcR gene in adults was detected at hypersalinity (42 ppt) and high temperature (35 °C) condition. The 5′-flanking region of TJ-EcR gene contains heat shock protein 70 response elements, implying that the environmental stressors may affect its expression via the stress-sensor. In addition, bisphenol A (100 µg/L) repressed TJ-EcR expression. Our results suggest that TJ-EcR could be a biomarker for the monitoring of the impact of environmental stressors in copepods.     

北京典型绿化树种幼苗光合特性对硬化地表的响应

以北京典型绿化树种油松(Pinus tabulaeformis)、白蜡(Fraxinus chinensis)和元宝枫(Acer truncatum)幼苗为试验材料,设置透水硬化地表、不透水硬化地表和对照3个水平的地表类型,通过监测地表温度、土壤含水量(20 cm)和光合参数等指标,分析3种树木叶片光合特性对硬化地表的响应特征。结果表明:城市硬化地表极显著地提高了地表温度(P0.01),表现为透水地表不透水地表对照;不透水地表极显著地降低了3种树木土壤含水量(P0.01);透水地表极显著地提高了油松和元宝枫土壤含水量(P0.01)。两种硬化地表下油松、元宝枫和不透水地表下白蜡幼苗叶片的净光合速率、蒸腾速率和气孔导度降低,株高和基径生长受到抑制。硬化地表对油松和元宝枫的影响主要源于硬化地表形成的高温胁迫,对白蜡的影响主要源于不透水地表形成的干旱胁迫。因此,在配置和种植城市绿化植物时,为缓解硬化地表对城市树木生长发育的不利影响,应选择适应高温和干旱的植物,并采取降温和灌溉措施来降低硬化地表的影响。     

乌鳢源杀鱼爱德华菌的分离鉴定及药敏特性研究

【背景】江苏省扬州市某乌鳢养殖场发生疾病,给养殖户造成了严重的经济损失。【目的】确定病因并筛选出敏感药物,为乌鳢相关疾病的防治提供参考。【方法】从患病乌鳢体内分离致病菌,并从形态、生理生化特征、16S rRNA和gyrB基因序列及特异性PCR检测等方面对分离菌株进行鉴定,同时开展人工感染试验分析其致病性,通过纸片扩散法进行药敏特性分析。【结果】从患病乌鳢体内分离获得优势菌株SHL,经形态特征、理化特性、16SrRNA和gyrB基因序列及特异性PCR检测鉴定为杀鱼爱德华菌。进一步人工感染试验证实其对乌鳢有较强的致病性,LD50为1.6×105 CFU/g,发病症状与自然发病症状相似。药敏试验结果显示,该菌株对青霉素、氯霉素、四环素等28种抗菌药物高度敏感,对红霉素中度敏感,对苯唑西啉、克拉霉素、万古霉素等6种药物耐药。【结论】引起江苏省扬州市某养殖场的乌鳢体表溃烂及死亡的病原菌为杀鱼爱德华菌,这是我国首次从淡水鱼类中检出致病性杀鱼爱德华菌,表明该菌的感染谱在扩大,需引起水产养殖领域的重视,在养殖过程中可根据药敏实验结果选用合适的国标渔药进行防治。     

加州鲈源鰤鱼诺卡氏菌的分离鉴定及致病性

[背景] 鰤鱼诺卡氏菌（Nocardia seriolae）是一种严重危害水产养殖业的病原菌,可引起以体表溃疡、出血及组织器官形成结节为特征的鱼类慢性肉芽肿疾病,目前尚无有效的防治方法。[目的] 明确引起安徽省临泉县某养殖场加州鲈（Micropterus salmonoides）结节病的病原菌,探讨其致病性,为该病的有效防治提供科学依据。[方法] 取肝脏结节病灶接种于TSB培养基分离优势细菌,利用表型检查结合分子生物学方法鉴定分离菌株。进一步通过检测分离菌株的毒力基因、测定其对加州鲈的半数致死量（LD₅₀）以及所感染加州鲈的组织病理学变化与组织载菌量,分析其致病性。[结果] 从病鱼体内分离到一株优势菌株NI,综合NI分离株的表型特性、16S rRNA基因序列与鰤鱼诺卡氏菌参考株相应序列的一致性以及特异性PCR扩增结果,确定其为鰤鱼诺卡氏菌。鰤鱼诺卡氏菌NI分离株携带毒力基因gapA、ibeA和mip,人工回归感染后加州鲈出现与自然病例相似的症状,其对加州鲈的LD₅₀为2.58×10⁶ CFU/尾。组织病理学观察到头肾、心脏、肝脏、胃和脾脏均出现慢性肉芽肿病变,肠管肌层疏松、肠绒毛脱落,肌肉组织中肌纤维疏松、间隙增宽。qPCR检测结果显示,组织中鰤鱼诺卡氏菌载量由高到低依次为头肾、心、肝、胃、脾、肠和肌肉。[结论] 鰤鱼诺卡氏菌是引起此次加州鲈结节病的病原菌,对该菌致病性的研究为加州鲈诺卡氏菌病的防控提供了理论依据。