Asexual multiplication of tetrathyridia of Mesocestoides corti in Crotalus viridis viridis

Prairie rattlesnakes, Crotalus viridis viridis Rafinesque, 1818, were infected with tetrathyridia of Mesocestoides corti Hoeppli, 1925, by means of oral intubation. Snakes were maintained at constant temperatures of either 25, 30, or 35 C for 4-24 wk. Postmortem examinations revealed the first experimental evidence of asexual multiplication of tetrathyridia in a laboratory-infected ectothermic host. Time and temperature were statistically significant with P values of less than 0.05.     

A new host and locality record for Mesocestoides sp. tetrathyridia (Cestoidea: Cyclophyllidea), with a summary of the genus from snakes of the world

A new host and distribution record is reported for tetrathyridia of Mesocestoides sp. One of 5 (20%) Namib tiger snakes, Telescopus beetzi, from South Africa was infected. Numerous tetrathyridia were found encapsulated in mesentery attached to the small intestine. Morphological examination of tetrathyridia revealed absence of buds, multiple scoleces, or any other evidence of asexual proliferation. A summary of the snakes of the world reported as hosts of tetrathyridia of Mesocestoides sp. is presented.     

Factors influencing the establishment of Mesocestoides corti in mice following oral inoculation of tetrathyridia

Factors which influence the establishment of tetrathyridia of Mesocestoides corti in mice following inoculation per os were examined. Only a proportion of the tetrathyridia penetrate the gut wall and gain access to the peritoneal cavity and liver, and most of these penetrate through the wall of the small intestine. It appears that tetrathyridia must attach to the intestinal mucosa and commence penetration immediately or they pass into the large intestine and are voided. Establishment was not influenced by strain, sex or age of host. However, the temperature at which tetrathyridia were maintained before inoculation influence their ability to penetrate the intestinal wall. Additionally it appears that tetrathyridia have to undergo a morphological change before or during, this penetration phase.     

Gene transfer system for Rhodopseudomonas viridis.

A gene transfer system for Rhodopseudomonas viridis was established which uses conjugation with Escherichia coli S17-I as the donor and mobilizable plasmids as vectors. Initially, plasmids of the incompatibility group P1 (pRK290 and pRK404) were used. The more effective shuttle vectors between E. coli and R. viridis, pKV1 and pKVS1, were derived from plasmid pBR322 and showed the highest conjugation frequency (10(-2] thus far demonstrated in purple bacteria. It was also demonstrated that Rhizobium meliloti can be used as a donor for conjugation with R. viridis. From a genomic cosmid library of R. viridis constructed in the vector pHC79, clones that coded for subunits H (puh operon), L, M and cytochrome c (puf operon) of the photosynthetic reaction center were isolated and characterized. For linkage of the two operons on the genome, cosmids that overlapped with the operon-carrying clones were identified. The relative positions of the two operons could not be determined, but the operons must be more than 100 kilobase pairs apart. Thus, the genomic organization of the reaction center in R. viridis is different from that of Rhodobacter capsulatus, for which a distance of about 39 kilobase pairs was determined. From a spontaneous mutant of R. viridis that is resistant to the herbicide terbutryn, the puf operon was cloned in pKVS1 and transferred by conjugation into R. viridis wild-type cells. The resulting exconjugants were resistant to the herbicide, which demonstrated that the puf operon on pKVS1 constructions was functionally expressed in R. viridis.     

Effect of bromocriptine on the larval skin of the green toad, Bufo viridis viridis leurenti

In the premetamorphic larval green toad, B. viridis viridis, as in other anurans, the skin is made up of a fibrous dermis and an epidermis of stratified epithelium. The effects of bromocriptine, an antiprolactin drug, on the premetamorphic skin of B. viridis viridis was examined. Bromocriptine, dissolved in rearing water at four different concentrations, induced a number of changes in the skin of treated tadpoles. In rough sequence of appearance, these changes include: retraction ofthe melanocyte dendrites, synchronous burst ofthe apical vesicles of the superficial epithelial cells, gradual disappearance of the melanosomes from the epithelial cells and widening of the intercellular spaces. In addition, macrophages appeared in the superficial dermis amongst the retracted melanocytes. White crystals were observed on the skin surface and similar crystals were ingested by the macrophages. Prolonged treatment with bromocriptine resulted in hypertrophy and extraction of some epidermal cells. Deep melanocytes of the mesenteries were not affected by bromocriptine-treatment indicating that the drug did not penetrate deep into the tadpole tissue. Whether the macrophages observed in the dermis were recruited from deeper tissues or were converted melanocytes is another issue in need of study.     

Occurrence of tetrathyridia of Mesocestoides sp. (Cestoidea: Cyclophyllidea) in North American anurans (Amphibia)

A new host and geographic locality record is reported for tetrathyridia of Mesocestoides sp. in two species of ranid frogs (Rana berlandieri and R. pipiens) from Texas and New York, respectively. Tetrathyridia were found encapsulated in liver and mesenteries of the hosts. Morphological examination and experimental inoculation of these tetrathyridia into mice demonstrated the absence of capacity for asexual proliferation. Overall prevalence of infection was low in anurans from Arkansas, Texas and New York, but intensities can be generally high. In addition, a summary of frogs and toads from North America reported as hosts of tetrathyridia of Mesocestoides sp. is presented.     

Isolation of new microsatellite loci from the Green Lizard (Lacerta viridis viridis)

Twelve new microsatellite loci were isolated from the Green Lizard (Lacerta viridis viridis). Primers for 28 loci were designed and 18 of these loci amplified well for 10 individuals of four populations. Twelve of these loci were further characterized for a population in Hungary. The results document the suitability of these identified loci for the characterization of the genetic diversity of the endangered species L. viridis viridis.     

一种狗尾草病原真菌的鉴定及菌株致病性研究

经形态学鉴定和rDNA ITS序列分析,16株分离自北京、河北、河南发病狗尾草的菌株、2株分别分离自河南发病虎尾草、牛筋草的菌株和1株分离自青海发病野燕麦的菌株被鉴定为狗尾草平脐蠕孢Bipolaris setariae。接种试验表明,来自狗尾草的菌株比来自其他寄主植物的菌株对狗尾草致病性强,分离自野燕麦的菌株对狗尾草无致病性,分离自不同地区不同样品狗尾草的菌株其致病性有显著差异。菌株NY1对狗尾草有很强致病性,接种后5d植株叶片即全部呈枯死状,接种后7d整个植株枯萎死亡。菌株NY1对马唐和虎尾草也有很强致病性,但对于大多数供试栽培植物致病性很弱或无致病性。因此,B. setariae NY1菌株具有进一步开发成为狗尾草、马唐和虎尾草等杂草的生物除草剂的潜力。     

Early post-larval development of the endoparasitic platyhelminth Mesocestoides corti: trypsin provokes reversible tegumental damage leading to serum-induced cell proliferation and growth

Mesocestoides corti is a suitable in vitro model for studying the development of human endoparasitic platyhelminthes. Treatment with trypsin, supplemented with fetal bovine serum (FBS), induces M. corti development from larvae (tetrathyridia) to segmented adult worm; however, the role of this protease and of FBS in post-larval development induction remains unknown. To characterize the participation of trypsin enzymatic activity and of FBS in the induction of tetrathyridia growth and development, both stimuli were added to the larvae either together or sequentially. Additionally, specific inhibition of trypsin activity was also monitored. Finally, the effect of the enzyme on the parasite tegument as well as the proliferative activity and location of proliferating cells after induction of tetrathyridia development were also studied. We conclude that trypsin-induced tetrathyridia development to adult worm is FBS-dependent and that the effect of serum factors is dependent upon a previous trypsin-induced reversible damage to the larva tegument. In dividing and non-dividing tetrathyridia, proliferative activity of cells is mainly located within the apical massif in the anterior region and nerve cords of larvae, respectively. In tetrathyridia stimulated to develop to adult worms, an intense proliferative activity is evident along the nerve cords. Our results suggest that in natural infections the tetrathyridia tegument is temporally made permeable to growth factors by proteolytic enzyme activity in the intestine juice of the definitive host, thus leading to development to adult worms.     

Assignment of the aromatic 1H-NMR resonances of myotoxin a isolated from the venom of Crotalus viridis viridis

The 400 MHz 1H-NMR spectrum of myotoxin a from the venom of Crotalus viridis viridis is described. The identification of spin systems in the aromatic region corresponding to the six aromatic residues of myotoxin a was completed using both one- and two-dimensional NMR spectroscopy and the pH dependence of chemical shifts. Assignments of these spin systems to specific residues was possible for the singly occurring amino acids Tyr-1 and Phe-12. Resonances from Tyr-1, His-5 and His-10 were shifted significantly from their random coil values in a pH-dependent manner. These shift perturbations were deemed evidence of a helical arrangement of the amino terminal region which placed these residues in close proximity to each other.     

A new small myotoxin from the venom of the prairie rattlesnake (Crotalus viridis viridis)

Fast atom bombardment (FAB) mass spectrometry was used to identify a new small myotoxin from the venom of the prairie rattlesnake (Crotalus viridis viridis). FAB mass spectrometry and Edman degradation were used to characterize its structure. This toxin is similar to myotoxin I from C. v. concolor, except that it possesses an additional. C-terminal asparaginyl-alanine. At 45 residues it is the longest known myotoxin a homolog. A myotoxin of 43 residues, identical to myotoxin I from C. v. concolor, was also found. To date no other species has been shown to produce more than one length of myotoxin. The present paper documents 42-, 43-, and 45-residue myotoxins from the venom of a single animal.     

Maternal transmission of asexually proliferative Mesocestoides corti tetrathyridia (Cestoda) in mice

Asexually proliferative Mesocestoides corti tetrathyridia were studied to test the hypothesis of in utero transmission in mice and define more clearly the path of transmammary transmission. In utero transmission was not observed in 132 fetuses (22 litters) taken by caesarean section from infected mothers. However, 19 of these mothers had tetrathyridia in their mammary glands at the time of operation, nine had worms in the uterine lumen, and one had a single worm in the maternal blood space of a placenta. No tetrathyridia were found in amniotic cavities. No infection was found in 32 young (7 litters) examined immediately after birth to infected mothers, but before nursing. No infection was found in 30 young (5 litters) removed from infected mothers before nursing and raised by uninfected fosters. Of 29 uninfected young (5 litters) allowed to nurse on infected mothers, 18 became infected. Whole mounts and sections of infected mammary glands showed proliferating tetrathyridia free in larger milk ducts and free and encapsulated in mammary parenchyma. These data suggest that maternal transmission of M. corti tetrathyridia in mice occurs primarily or perhaps exclusively by the transmammary route.     

Urea tolerance and osmoregulation in Bufo viridis and Rana ridibunda

1. The tolerance and adaptation to urea solutions by terrestrial green toads (Bufo viridis) and semi-aquatic frogs (Rana Ridibunda) were studied. 2. the green toad showed tolerance to urea solution of 800 mM and the frogs showed tolerance only to about 400 mM urea solution. 3. The plasma concentrations of both species was hyperosmotic to the external medium in all the different urea solutions. 4. Blood osmolality, urea, Na+ and Cl- concentrations of B. viridis were always higher than in R. ridibunda. 5. The urea concentration in muscle of R. ridibunda was higher than the urea concentration in muscle of B. viridis. 6. The muscle tissue weight loss of B. viridis was significantly lower than R. ridibunda.     

葡萄糖诱导绿色杜氏藻转录组及相关通路差异分析

绿色杜氏藻是研究耐盐机理的模式绿藻．葡萄糖不仅是营养物质,而且还是信号物质．目前,对绿色杜氏藻转录组、糖处理后差异表达基因和β-胡萝卜素生物合成途径关键基因表达还不清楚．本研究通过Illumina HiSeqTM 2000高通量测序,获得葡萄糖处理和未处理绿色杜氏藻转录组信息．利用P value值和差异倍数对样本进行差异表达分析,共111条转录本存在差异表达,3条为上调转录本,108条为下调转录本．利用RT-qPCR检验差异表达分析的准确性. 结果表明,转录本表达结果与转录组分析结果一致．GO功能富集结果表明,71条下调转录本与代谢相关,占所有下调转录本的65.74%．KEGG富集分析结果表明,21条KEGG通路含89条下调转录本,14条通路与代谢相关．代谢中通路最多的为能量代谢（6条）,含63条下调转录本．能量代谢中与光合作用相关的下调转录本最多,为29条．通过分析找到2条与β-胡萝卜素生物合成相关通路（MVA/MEP途径及β-胡萝卜素合成途径）,并发现通路的关键基因hmgs、dxs、dxr、psy、pds、chyb,对其进行差异表达分析,均不存在差异表达．研究表明,葡萄糖抑制了绿色杜氏藻光合作用,代谢受阻,但未影响β-胡萝卜素生物合成相关通路及关键基因．     

The complete mitochondrial genome of the green lizard Lacerta viridis viridis (Reptilia: Lacertidae) and its phylogenetic position within squamate reptiles

For the first time the complete mitochondrial genome was sequenced for a member of Lacertidae. Lacerta viridis viridis was sequenced in order to compare the phylogenetic relationships of this family to other reptilian lineages. Using the long-polymerase chain reaction (long PCR) we characterized a mitochondrial genome, 17,156 bp long showing a typical vertebrate pattern with 13 protein coding genes, 22 transfer RNAs (tRNA), two ribosomal RNAs (rRNA) and one major noncoding region. The noncoding region of L. v. viridis was characterized by a conspicuous 35 bp tandem repeat at its 5' terminus. A phylogenetic study including all currently available squamate mitochondrial sequences demonstrates the position of Lacertidae within a monophyletic squamate group. We obtained a narrow relationship of Lacertidae to Scincidae, Iguanidae, Varanidae, Anguidae, and Cordylidae. Although, the internal relationships within this group yielded only a weak resolution and low bootstrap support, the revealed relationships were more congruent with morphological studies than with recent molecular analyses.     

Improved medium for recovery and enumeration of the farmer's lung organism, Saccharomonospora viridis

A new medium, which we propose to call R8, was developed for the isolation and enumeration of the thermophilic actinomycete, Saccharomonospora viridis. This organism has been implicated in a range of hypersensitivity pneumonitides, including farmer's lung, but is generally isolated in small numbers from contaminated environments. Recovery of S. viridis from moldy hay and mushroom compost on R8 medium was compared with recovery on conventional media. S. viridis was isolated from both substrates but in highest numbers and most consistently on the R8 medium. The selectivity of this medium was best observed when the sedimentation chamber method was used for hay samples. Here S. viridis accounted for up to 80% of the total number of actinomycetes recovered on R8 and could not be recovered on rifampin selective medium under the same conditions. R8 was also found to be an efficient recovery medium for a range of thermophilic actinomycetes from mushroom compost and for another allergenic species, Faenia rectivirgula, from moldy hay. Contamination of isolation plates by thermophilic bacilli was reduced on R8 compared with the activity on half-strength tryptone soy agar, supplemented with 0.2% casein hydrolysate, and this, together with specific improvements in S. viridis growth, accounts for the selective effect. It is possible that the occurrence of S. viridis and its role as a causative agent of hypersensitivity pnuemonitis have been underestimated by the use of suboptimal recovery protocols. It is hoped that use of R8 in conjunction with dilution plate techniques will generate information on the ecology of S. viridis and contribute to health risk assessment studies.     

Improved medium for recovery and enumeration of the farmer's lung organism, Saccharomonospora viridis.

A new medium, which we propose to call R8, was developed for the isolation and enumeration of the thermophilic actinomycete, Saccharomonospora viridis. This organism has been implicated in a range of hypersensitivity pneumonitides, including farmer's lung, but is generally isolated in small numbers from contaminated environments. Recovery of S. viridis from moldy hay and mushroom compost on R8 medium was compared with recovery on conventional media. S. viridis was isolated from both substrates but in highest numbers and most consistently on the R8 medium. The selectivity of this medium was best observed when the sedimentation chamber method was used for hay samples. Here S. viridis accounted for up to 80% of the total number of actinomycetes recovered on R8 and could not be recovered on rifampin selective medium under the same conditions. R8 was also found to be an efficient recovery medium for a range of thermophilic actinomycetes from mushroom compost and for another allergenic species, Faenia rectivirgula, from moldy hay. Contamination of isolation plates by thermophilic bacilli was reduced on R8 compared with the activity on half-strength tryptone soy agar, supplemented with 0.2% casein hydrolysate, and this, together with specific improvements in S. viridis growth, accounts for the selective effect. It is possible that the occurrence of S. viridis and its role as a causative agent of hypersensitivity pnuemonitis have been underestimated by the use of suboptimal recovery protocols. It is hoped that use of R8 in conjunction with dilution plate techniques will generate information on the ecology of S. viridis and contribute to health risk assessment studies.     

Population dynamics and diapause response of the springtail pest Sminthurus viridis (Collembola: Sminthuridae) in southeastern Australia

The springtail, Sminthurus viridis (L.) (Collembola: Sminthuridae), is an important agricultural pest across southern Australia. We investigated the seasonal abundance patterns and summer diapause response of S. viridis in southeastern Australia by using field and shadehouse (a greenhouse that offers seedlings shade) experiments. Seasonal activity patterns of S. viridis were largely consistent with previous studies, with the pest active from autumn to spring. In addition, the timing and pattern of the summer-diapausing egg stage was established, with multiple generations probably producing diapause eggs. A strong relationship between soil moisture and temperature with autumn emergence also was observed. These results suggest that S. viridis autumn pest pressure can be predicted and indicate that late-season spraying strategies currently used for a sympatric agricultural pest are unlikely to be as effective against S. viridis.     

绿色巴夫藻和四列藻种间竞争机制研究

种间竞争是生物普遍存在的一种生命现象 ,可以利用来调控生物的生长和数量变化 ,进行有害生物的防治。藻类植物间也存在明显的竞争现象。Ｈｏｌｍ[8] 报道了硅藻和微囊藻种间的相互作用。Ｈｅｇａｒｔｙ[9] 研究了光强和Ｎ、Ｐ比对棕囊藻和 5种硅藻间竞争的影响。Ｋｕｗａｔａ[10 ] 研究了铵盐对绿藻和蓝藻间竞争作用的影响。Ｄａｖｉｓ[11] 研究了海藻和海草间的竞争。Ｐｉａｚｚｉ[12 ] 研究了两种海洋绿藻生长的竞争情况。刘世枚[1] 研究了两种绿藻种群间的相互作用。陈德辉等[2 ] 对微囊藻和栅藻共培养进行了研究并计算了竞争参数。。绿…     

Parasites of prairie rattlesnakes (Crotalus viridis viridis) and gopher snakes (Pituophis melanoleucus sayi) from the eastern high plains of New Mexico

Three prairie rattlesnakes (Crotalus viridis viridis) and two gopher snakes (Pituophis melanoleucus sayi) from the eastern high plains of New Mexico (USA) were examined for parasites. One cestode (Oochoristica osheroffi), and two nematode (Kalicephalus inermis and Physoloptera retusa) species were recovered from two infected rattlesnakes. One female gopher snake was infected with two nematode (K. inermis and Rhabdias spp.) and one mite (Entonyssus halli) species.