Morphology and ultrastructure of the Malpighian tubules of the Chilean common tarantula (Araneae: Theraphosidae).

Relatively little is known about the morphology and ultrastructure of the Malpighian tubules of spiders (Arachnida: Araneae). Our study represents the first investigation of the Malpighian tubules of a theraphosid spider and is the only study to examine the living Malpighian tubules using confocal laser scanning microscopy. In theraphosid spiders, the Malpighian tubules originate from the stercoral pocket in the posterior portion of the opisthosoma and extend forward toward the prosoma in a dendritic pattern. There are three distinct segments (initial, main, and terminal), all dark brown in appearance. Each segment has distinctive ultrastructural features. Both the terminal and the main segment appear to be composed of at least two cell types with finger-like cytoplasmic protrusions associated with one of these types. The terminal segment, which is most proximal to the stercoral pocket, is the largest in diameter. It is composed of large, cuboidal cells containing many mitochondria and lipid inclusions. The main segment is intermediate in diameter with many mitochondria and secretory vesicles present. The initial segment is relatively thin in comparison to the other segments and is intimately associated with the digestive gland. The cells of the initial segment contain very little cytoplasm, fewer mitochondria, secretory vesicles, and prominent inclusions.     

Ultrastructural evidence for paracellular fluid flow in the Malpighian tubules of a larval mayfly

The ultrastructure of the Malpighian tubules of larvae of the Mayfly Ecdyonurus dispar (Ephemeroptera) is described. There are about 60 tubules, which consist of four distinct regions. The most proximal section (region I) appears to be responsible for fluid secretion. A unique feature is the presence of channels leading off the main lumen, which end close to the basal border of the cells. Microvilli are confined to these channels in region I. Region II is a short spiral region, the cells of which possess long basal folds and associated mitochondria. Region III is a simple conducting tube leading to one of six collecting ducts (region IV) arranged radially around the gut. In each collecting duct there are two cell types present. Type 2 cells are relatively simple, but give rise to numerous, long, microvilli-like projections. Type 1 cells possess long basal folds, and curious membrane whorls in the apical zone. Evidence is presented which suggest that water movements into region I takes place via the paracellular route. Region II is probably a reabsorptive region, but the function of region IV, based on ultrastructural evidence is more difficult to elucidate.     

Ultrastructural changes in the muscles,midgut, hemopoietic organ,imaginal discs and Malpighian tubules of the mosquito Aedes aegypti larvae infected by the fungus Coelomomyces stegomyiae

Fungi belonging to the genus Coelomomyces can infect mosquito larvae and develop within the larval hemocoel. To examine fungal development, Aedesaegypti larvae infected with Coelomomyces stegomyiae Keilin were fixed, embedded and sectioned for both light and electron microscopy. While fungal hyphae of C. stegomyiae did not invade cells other than the cuticular epithelial cells, they did penetrate a number of tissues including muscles, midgut, hemopoietic organ, imaginal discs, and Malpighian tubules. This revised version was published online in July 2006 with corrections to the Cover Date.     

庭疾灶螽中肠及马氏管结构

【目的】本研究旨在以庭疾灶螽Tachycines asynamorus为例探索驼螽消化系统和排泄系统在结构上与其生活环境的适应关系。【方法】运用解剖学方法、石蜡切片技术、冰冻切片技术及超薄切片技术对庭疾灶螽中肠及马氏管的结构进行研究。【结果】庭疾灶螽中肠向前延伸出3个胃盲囊包围着前胃。中肠上皮由再生细胞、柱状上皮细胞和内分泌细胞构成,具有典型的再生细胞龛;闭合型内分泌细胞紧贴在再生细胞龛的外围,基底区聚集大量的分泌颗粒。柱状上皮细胞内聚集有2类大的分泌颗粒：线团状颗粒和电子密度很高的球状颗粒;中肠管腔内有明显的围食膜结构,中肠基底部由基膜和肌肉层组成。马氏管着生在中后肠的交界处,从横切面看马氏管管壁具有3~5个细胞,细胞近管腔端部具有大量长微绒毛,细胞质内分布着电子致密的同心圆球晶体,基底膜内折形成膜迷路。【结论】庭疾灶螽中肠柱状上皮细胞的线团状颗粒由微丝包裹;内分泌细胞由再生细胞龛中的细胞分化而来,产生内分泌颗粒并将其排到血腔;中肠基膜发达,包含微丝与复合糖成分,基膜通过对中肠上皮细胞的支撑作用为肠道蠕动提供保障。庭疾灶螽马氏管细胞中可见大量颗粒和大量同心圆球晶体,推测可能是一种储存排泄。     

The effects of endogenous diuretic and antidiuretic peptides and their second messengers in the Malpighian tubules of Tenebrio molitor: an electrophysiological study

The Malpighian tubules of Tenebrio molitor provide a model system for interpreting the actions of endogenous diuretic and antidiuretic peptides. The effects of diuretic (Tenmo-DH(37)) and antidiuretic (Tenmo-ADFa) peptides and their respective second messengers (cyclic AMP and cyclic GMP) on basolateral (V(bl)) and transepithelial (V(te)) potentials of Tenebrio Malpighian tubules were determined using conventional microelectrodes. In the presence of 6 mmol l(-1) Ba(2+), Tenmo-DH(37) (100 nmol l(-1)) reversibly hyperpolarized V(bl) and depolarized V(te). A similar response was seen with the addition of 1 mmol l(-1) cyclic AMP; however, the apical membrane potential (V(ap)) then showed a hyperpolarization, whereas a depolarization of V(ap) was observed with Tenmo-DH(37). Bafilomycin A(1) (5 micromol l(-1)) inhibited fluid secretion of stimulated tubules and reversed the hyperpolarization of V(bl) in response to Tenmo-DH(37). In response to 100 nmol l(-1) Tenmo-ADFa or 1 mmol l(-1) cyclic GMP, V(bl) and V(te) depolarized, although cyclic GMP affected membrane potentials somewhat differently by causing an initial hyperpolarization of V(bl) and V(te). In high [K(+)]-low [Na(+)] Ringer, 1 mmol l(-1) amiloride decreased fluid secretion rates, and depolarized both V(bl) and V(te). Amiloride significantly decreased luminal pH in paired experiments, indicating the presence of a K(+)/nH(+) exchanger in tubule cells of Tenebrio. The results suggest that the endogenous factors and their second messengers stimulate/inhibit fluid secretion by acting on the apical V-ATPase, basolateral K(+) transport, and possibly Cl(-) transport.     

The effect of infection with Dirofilaria immitis (dog heartworm) on fluid secretion rates in the Malpighian tubules of the mosquitoes Aedes taeniorhynchus and Anopheles quadrimaculatus

Dirofilaria immitis, the parasitic nematode causing the disease dog heartworm, is transmitted by female mosquitoes. During their development, larval nematodes reside in the cells of the Malpighian tubules of these mosquitoes for approx 13 days. We have examined the effect of the presence of these large intracellular parasites on the main physiological function of the Malpighian tubules, i.e. fluid secretion. Rates of fluid secretion were examined in vitro using both normal and infected tubules of the mosquito species Aedes taeniorhynchus and Anopheles quadrimaculatus. Tubules of A. quadrimaculatus show changes in trasport rate during the reproductive cycle. Those of A. taeniorhynchus do not. Infection with larvae of D. immitis had no effect on the rate of fluid secretion in tubules of A. quadrimaculatus. In A. taeniorhynchus by contrast, the tubules show decline in transport with time following infection. The reduction in transport capacity is proportional to the number of worms infecting the tubule. The present paper and separate ultrastructural studies demonstrate that parallel changes in microvillar ultrastructure and epithelial transport rates occur in response to infection by the parasite. In both species examined, the survival of the mosquitoes and their vector potential are determined by factors other than the transport capacities of the infected Malpighian tubules.     

Effects of bafilomycin A1 and amiloride on the apical potassium and proton gradients in Drosophila Malpighian tubules studied by X-ray microanalysis and microelectrode measurements

The intracellular distribution of potassium in Malpighian tubules from Drosophila larva was measured by electron probe X-ray microanalysis of freeze-dried cryosections. Application of amiloride alone to the haemolymph space had no effect on the intracellular potassium concentration in the region of intermediate cytoplasm (between the basal region of basal membrane infoldings and the apical brush border), whereas a potassium increase as well as a chloride increase was observed after simultaneous blocking of the potassium conductance of the basal membrane with barium. Injected bafilomycin and amiloride applied in the haemolymph caused an increase of the potassium content in the basal cytoplasm but not in the microvilli. In addition, the intracellular water portion was decreased by bafilomycin. pH measurements in isolated larval anterior tubules with proton-selective microelectrodes showed that bafilomycin added to the bathing solution caused a decrease in intracellular pH. Addition of amiloride had no significant effect on intracellular pH, but the pH of the luminal fluid was decreased within 1 min by 0.5 pH units. The amiloride-induced luminal pH decrease could be inhibited by the metabolic blocker KCN as well as by bafilomycin. Furthermore, removing potassium from the bathing saline caused a slow luminal acidification, which could be blocked by KCN. Our results support the hypothesis of a functionally coupled transport system in the apical membrane consisting of a bafilomycin-sensitive V-ATPase and a K⁺-dependent, amiloride-sensitive K⁺/H⁺ exchange system.Abbreviation C _a element concentration related to water - C _d element content related to dry weight - dw dry weight - DMSO dimethylsulphoxide - emf electromotive force - NBD-Cl 7-chloro-4-nitrobenz-2-oxa-1,3-diazole - NEM N-ethylmaleimide - NMDG⁺ N-methyl-d-glucamine - PD potential difference - pH_i intracellular pH value - pH_lu luminal pH value - pmf protonmotive force - SD standard deviation - SE standard error - STEM scanning transmission electron microscopy - V _a apical potential difference - V _b basal potential difference - V _t transepithelial potential difference     

Ultrastructural zonal heterogeneity of hepatocytes and mitochondria within the hepatic acinus during liver regeneration after partial hepatectomy

BACKGROUND INFORMATION: Partial hepatectomy (70%) induces cell proliferation until the original mass of the liver is restored. In the first 24 h after partial hepatectomy, drastic changes in the metabolism of the remaining liver have been shown to occur. To evaluate changes in hepatocyte ultrastructure within the hepatic acinus during the liver regenerative process, we investigated, by light and electron microscopy observations on specimens taken 0 h, 24 h and 96 h after partial hepatectomy, the hepatocyte structure and ultrastructure in the periportal and pericentral area of the hepatic acinus, with a particular emphasis on mitochondria ultrastructure. Moreover, some biochemical events that could affect the mitochondria ultrastructure and function were investigated. RESULTS: We found that, 24 h after partial hepatectomy, mitochondria with altered ultrastructure were preferentially localized in the periportal area. Periportal hepatocytes showed also an increase in the number of peroxisomes, free ribosomes, lysosomes and autophagosomes. Altered mitochondria showed swelling, an ultrastructural index of increased membrane permeability, a reduction in the number of cristae, and a rarefied, often vacuoled, matrix. Consistently, an increase in the mitochondrial oxidized/reduced glutathione ratio was found as well as calcium release from mitochondria in a manner inhibited by cyclosporin A. Interestingly, light and electron microscopy analysis showed that the hepatocytes in the periportal area were the cells with the major structural attributes to proliferate. At 96 h after partial hepatectomy, the preferential zonation of altered mitochondria was lost and the normal mitochondrial membrane permeability properties were restored. CONCLUSIONS: We suggest that 24 h after partial hepatectomy, a preferential zonation of altered mitochondria in the periportal hepatocytes could be involved in the changes of metabolic and functional heterogeneity of the hepatocytes within the hepatic acinus during the regenerative process.     

Structure of galactarate dehydratase,a new fold in an enolase involved in bacterial fitness after antibiotic treatment

Galactarate dehydratase (GarD) is the first enzyme in the galactarate/glucarate pathway and catalyzes the dehydration of galactarate to 3‐keto‐5‐dehydroxygalactarate. This protein is known to increase colonization fitness of intestinal pathogens in antibiotic‐treated mice and to promote bacterial survival during stress. The galactarate/glucarate pathway is widespread in bacteria, but not in humans, and thus could be a target to develop new inhibitors for use in combination therapy to combat antibiotic resistance. The structure of almost all the enzymes of the galactarate/glucarate pathway were solved previously, except for GarD, for which only the structure of the N‐terminal domain was determined previously. Herein, we report the first crystal structure of full‐length GarD solved using a seleno‐methoionine derivative revealing a new protein fold. The protein consists of three domains, each presenting a novel twist as compared to their distant homologs. GarD in the crystal structure forms dimers and each monomer consists of three domains. The N‐terminal domain is comprised of a β‐clip fold, connected to the second domain by a long unstructured linker. The second domain serves as a dimerization interface between two monomers. The C‐terminal domain forms an unusual variant of a Rossmann fold with a crossover and is built around a seven‐stranded parallel β‐sheet supported by nine α‐helices. A metal binding site in the C‐terminal domain is occupied by Ca²⁺. The activity of GarD was corroborated by the production of 5‐keto‐4‐deoxy‐D‐glucarate under reducing conditions and in the presence of iron. Thus, GarD is an unusual enolase with a novel protein fold never previously seen in this class of enzymes.     

Lead toxicity at different life stages of the giant prawn (Macrobrachium rosenbergii,de Man): considerations of osmoregulatory capacity and histological changes in adult gills

This study evaluated the acute toxicity of lead in different life stages of the freshwater prawn Macrobrachium rosenbergii, determined the effect of sublethal Pb concentrations on osmoregulatory capacity (OC), measured the Pb level in gills, and investigated the effect of Pb on the structure of the gills of adult prawns. The 24-, 48-, and 96-h LC₅₀ values for Pb to M. rosenbergii increased progressively with increasing life stage, from post-larvae (PL), juvenile to adult. The 24- and 48-h LC₅₀ values for post-larvae ranged from 0.01- to 0.09-mg Pb/L. The 24-, 48-, and 96-h LC₅₀ values for Pb were lower at 12 ppt than those at 0 ppt for either the juveniles or the adults. At 12 ppt, the 96-h LC₅₀ values in PL11, juvenile and adult were 0.47-, 0.58-, and 2.03-mg Pb/L, respectively. Meanwhile, at 12 ppt, the 96-h LC₅₀ values in PL11, juvenile and adult were 0.63-, 4.44-, and 7.98-mg Pb/L, respectively. In adults, the OC values of controls and prawns exposed to 2- and 4-mg Pb/L at 0 ppt were not significantly different. The OC of prawns exposed to and 2-mg Pb/L at 12 ppt increased by 72 and 109% from the OC of the control prawns. At media 12 ppt, the OC value of prawns exposed to 1-mg Pb/L was significantly different from that of prawns exposed to 2-mg Pb/L. The concentrations of Pb in gill tissues increased significantly in Pb exposed prawns both at 0 and 12 ppt. The level of Pb in gills of prawns exposed to 2-mg Pb/L at 12 ppt was not significantly different from those exposed at 0 ppt. The severe toxic actions of Pb were noted in gills of prawns exposed in media 12 ppt. Hyperplasia and necrosis were observed in gill lamellae, resulting in abnormal gill tips after Pb exposure at media 12 ppt. Since the effect of Pb is more pronounced in higher salinity (12 ppt) than in freshwater (0 ppt) it is clear that aquaculture of M. rosenbergii should be conducted in freshwater ponds.     

Characterization of transepithelial transport of salicylate by the Malpighian tubules of Drosophila melanogaster and the effects of changes in fluid secretion rate

Abstract.  A radioisotope tracer technique is used to study mechanisms and regulation of transepithelial transport of the plant allelochemical salicylate by the Malpighian tubules of Drosophila melanogaster . Transepithelial transport of salicylate is nearly abolished in Na⁺-free saline, and inhibited by ouabain, low K⁺ or K⁺-free bathing saline. In addition, the carboxylates probenecid, unlabelled salicylate, fluorescein, and p -aminohippuric acid (PAH) significantly inhibit transepithelial transport of salicylate. The sulphonates taurocholate and phenol red also inhibit transepithelial transport of salicylate, whereas amaranth has no effect. Stimulation of fluid secretion by cAMP, cGMP or leucokinin I increases transepithelial transport of salicylate, particularly when the concentration of salicylate in the bathing saline is high. The correlation between the fluid secretion rate and transepithelial transport of salicylate shows that 64% of the changes in salicylate transport can be explained on the basis of changes in fluid secretion rate. The results show that naturally-occurring plant secondary metabolite salicylate is transported into the lumen of the Mapighian tubules of D. melanogaster by a mechanism similar to that previously described for the prototypical organic anions PAH and fluorescein. In addition, the transepithelial transport of salicylate increases in response to increases in fluid secretion rate.     

Biochemical activity and multiple locations of particulate guanylate cyclase in Rhyacophila dorsalis acutidens (Insecta: Trichoptera) provide insights into the cGMP signalling pathway in Malpighian tubules

In insect renal physiology, cGMP and cAMP have important regulatory roles. In Drosophila melanogaster, considered a good model for molecular physiology studies, and in other insects, cGMP and cAMP act as signalling molecules in the Malpighian tubules (MTs).However, many questions related to cyclic nucleotide functions are unsolved in principal cells (PC) and stellate cells (SC), the two cell types that compose the MT. In PC, despite the large body of information available on soluble guanylate cyclase (sGC) in the cGMP pathway, the functional circuit of particulate guanylate cyclase (pGC) remains obscure. In SC, on the other side, the synthesis and physiological role of the cGMP are still unknown. Our biochemical data regarding the presence of cyclic nucleotides in the MTs of Rhyacophila dorsalis acutidens revealed a cGMP level above the 50%, in comparison with the cAMP. The specific activity values for the membrane-bound guanylate cyclase were also recorded, implying that, besides the sGC, pGC is a physiologically relevant source of cGMP in MTs. Cytochemical studies showed ultrastructurally that there was a great deal of pGC on the basolateral membranes of both the principal and stellate cells. In addition, pGC was also detected in the contact zone between the two cell types and in the apical microvillar region of the stellate cells bordering the tubule lumen. The pGC signal is so well represented in PC and, unexpectedly in SC of MTs, that it is possible to hypothesize the existence of still uncharacterized physiological processes regulated by the pGC-cGMP system.     

High-performance liquid chromatographic methods for the determination of a new carbapenem antibiotic, L-749,345, in human plasma and urine

A column-switching, reversed-phase high-performance liquid chromatographic (HPLC) method for the determination of a new carbapenem antibiotic assay using ultraviolet detection has been developed for a new carbapenem antibiotic L-749,345 in human plasma and urine. A plasma sample is centrifuged and then injected onto an extraction column using 25 mM phosphate buffer, pH 6.5. After 3 min, using a column-switching valve, the analyte is back-flushed with 10.5% methanol–phosphate buffer for 3 min onto a Hypersil 5 μm C₁₈ BDS 100×4.6 mm analytical column and then detected by absorbance at 300 nm. The sample preparation and HPLC conditions for the urine assay are similar, except for a longer analytical column 150×4.6 mm. The plasma assay is specific and linear from 0.125 to 50 μg/ml; the urine assay is linear from 1.25 to 100 μg/ml.     

海州湾近岸海域的微生物群落结构和抗生素抗性基因时空变化

【背景】近岸海域抗生素抗性基因(antibiotic resistance genes, ARGs)的污染和累积将直接影响海产品质量和安全,海州湾作为江苏省的四大渔场之一,是江苏渔业发展的主要载体,有多条大小河流注入,沿岸为重要农业区,对公众健康产生重大影响。【目的】对海州湾夏秋季的水样及沉积物展开微生物及ARGs检测。【方法】基于宏基因组测序技术开展海州湾夏秋两季近岸6个站点中水体和沉积物中ARGs种类和相对丰度以及微生物群落的组成研究。【结果】变形菌门(Proteobacteria)和放线菌门(Actinobacteria)是夏秋季度两种介质中最优势的门类,水样中优势的科级细菌为红细菌科(Rhodobacteraceae),沉积物样品中为脱硫杆菌科(Desulfobacteraceae);夏季水样中的ARGs相对丰度要明显高于秋季,但沉积物中不同季节的ARGs相对丰度未表现出明显的变化趋势;在水样中主要门级微生物群落的抗性机制主要是抗生素靶位替换和抗生素靶位保护,沉积物样品则以抗生素灭活机制为主,而主要科级微生物群落的抗性机制更加多样;冗余分析(redundancyanalysis...     

Phenotypic changes and reduced genetic diversity have accompanied the rapid decline of the garden tiger moth (Arctia caja) in the U.K.

Abstract 1. Previous studies have quantified the recent decline of numerous Lepidopteran species in the U.K., including the garden tiger moth (Arctia caja), in which abundance has decreased by 85% over the past 30 years. At the same time that overall numbers have been falling, the distribution of abundance of this species has been moving northwards. In this study, morphological and genetic data were used to investigate the possibility that these changes in abundance and distribution have been accompanied by microevolutionary changes. 2. A comparison of wing size and shape in current and historical moth samples revealed that wing shape has altered significantly over the past century, resulting in longer, narrower hindwings and narrower forewings for a given forewing length. Habitat fragmentation and increased suitability of northerly sites provide a plausible explanation for the selection of increasingly dispersive individuals. 3. Mitochondrial DNA revealed no phylogeographic structuring either before or after the population decline. However, a comparison of mtDNA haplotypes from current and museum specimens indicated that the recent population decline across the U.K. has been accompanied by a significant loss of genetic diversity. 4. The changes in wing shape suggest recent adaptation to environmental change, whereas a loss of genetic diversity may limit the ability of garden tiger moths to adapt to future environmental change.     

一对紫绀型先天性心脏病患儿肠道菌群动态变化及阴沟肠杆菌耐药性研究

[目的]观察一对紫绀型先天性心脏病双胞胎新生儿在接受手术及抗生素治疗前后的肠道菌群的动态变化,探究患儿肠道内阴沟肠杆菌在治疗期间耐药性如何发生改变。[方法]通过采集患儿在不同治疗阶段的粪便样本,进行培养组学和16S rRNA基因测序分析。同时,从双胞胎哥哥不同治疗阶段的粪便样本中分离得到10株阴沟肠杆菌进行体外药敏试验。[结果]培养组学结果显示使用抗生素导致术前粪便样本丰富度降低,仅可分离获得肠球菌属、不动杆菌属和肠杆菌属;长期医院环境暴露后,能分离出多种条件致病菌属,包括肠杆菌属、克雷伯菌属、不动杆菌属和假单胞菌属;母乳喂养4个月后,粪便菌群丰富度增加、构成发生改变,可分离获得乳杆菌属和双歧杆菌属等有益细菌。16S rRNA基因测序结果显示,在多种因素影响下不同治疗时间点α多样性指数和相对丰度最高的菌属不同。细菌耐药实验结果显示,治疗期间阴沟肠杆菌对哌拉西林逐渐产生耐药性。[结论]接受抗生素治疗、院内环境暴露和母乳喂养共同影响患儿治疗和康复期间的肠道菌群组成;在不同治疗阶段仅使用一种抗生素,也会导致阴沟肠杆菌对不同药物的耐药性发生改变。     

Sandflies of the Phlebotomus perniciosus complex: mitochondrial introgression and a new sibling species of P. longicuspis in the Moroccan Rif

The bloodsucking adult females of Phlebotomus perniciosus Newstead and P. longicuspis Nitzulescu (Diptera: Psychodidae) are important vectors of the protozoan Leishmania infantum Nicolle (Kinetoplastida: Trypanosomatidae) in western Mediterranean countries. The species status of the two phlebotomine sandflies was assessed, along with the epidemiological implications. Individual sandflies from three Moroccan Rif populations were characterized morphologically, isoenzymatically (by the isoelectrofocusing of alleles at the polymorphic enzyme loci of HK, GPI and PGM), and by comparative DNA sequence analysis of a fragment of mitochondrial Cytochrome b (mtDNA). By reference to the character profiles of specimens from other locations, including southern Spain and the type-locality countries, the Moroccan flies were placed in three lineages: first, the lineage of P. perniciosus, which contained two mtDNA sublineages, one (pnt) widely distributed and associated with the morphology of the male types from Malta, and the other (pna) associated with a P. longicuspis-like male morphology; second, the lineage of P. longicuspis sensu stricto, including typical forms from Tunisia; and third, a new sibling species of P. longicuspis. The mtDNA sublineage (pnt) of typical P. perniciosus was also found in some P. longicuspis from Morocco, indicating interspecific hybridization. The typical race of P. perniciosus occurs in Italy as well as in Malta, Tunisia and Morocco. It is replaced in southern Spain by the Iberian race (with the pni mtDNA sublineage). The discovery of interspecific gene introgression and a new sibling species mean that previous records of the two morphospecies do not necessarily reflect their true vectorial roles or geographical and ecological distributions.     

Temporal changes of symbiont density and host fitness after rifampicin treatment in a whitefly of the Bemisia tabaci species complex

Microbial symbionts are essential or important partners to phloem‐feeding insects. Antibiotics have been used to selectively eliminate symbionts from their host insects and establish host lines with or without certain symbionts for investigating functions of the symbionts. In this study, using the antibiotic rifampicin we attempted to selectively eliminate certain symbionts from a population of the Middle East‐Asia Minor 1 whitefly of the Bemisia tabaci species complex, which harbors the primary symbiont “Candidatus Portiera aleyrodidarum” and two secondary symbionts “Candidatus Hamiltonella defensa” and Rickettsia. Neither the primary nor the secondary symbionts were completely depleted in the adults (F0) that fed for 48 h on a diet treated with rifampicin at concentrations of 1–100 μg/mL. However, both the primary and secondary symbionts were nearly completely depleted in the offspring (F1) of the rifampicin‐treated adults. Although the F1 adults produced some eggs (F2), most of the eggs failed to hatch and none of them reached the second instar, and consequently the rifampicin‐treated whitefly colony vanished at the F2 generation. Interestingly, quantitative polymerase chain reaction assays showed that in the rifampicin‐treated whiteflies, the density of the primary symbiont was reduced at an obviously slower pace than the secondary symbionts. Mating experiments between rifampicin‐treated and untreated adults demonstrated that the negative effects of rifampicin on host fitness were expressed when the females were treated by the antibiotic, and whether males were treated or not by the antibiotic had little contribution to the negative effects. These observations indicate that with this whitefly population it is not feasible to selectively eliminate the secondary symbionts using rifampicin without affecting the primary symbiont and establish host lines for experimental studies. However, the extinction of the whitefly colony at the second generation after rifampicin treatment indicates the potential of the antibiotic as a control agent of the whitefly pest.     

Morphofunctional changes of the digestive gland in the bivalve mollusk Crenomytilus grayanus (Dunker, 1853) in normal conditions and after parasitic invasion by trematodes

Specifics of the digestive cycle were studied in normal individuals of Gray’s mussel Crenomytilus grayanus and in those infested by trematodes from an unpolluted area of the sublittoral zone of Peter the Great Bay in May (during the day), June, July, and August. Four types of tubules corresponding to four phases of the digestive cycle were identified, and three varieties of destructing tubules were distinguished. It was shown that normally phases of absorption and digestion prevailed in the digestive gland. The portion of absorptive tubules (type II) was 95% in spring and 79% in summer. The total portion of destructing and restoring tubules (types III and IV) did not exceed 10% in spring and 20% in summer, and portion of tubules at the initial condition (type I) was 4% in spring and 2% in summer. During the day, insignificant variations in the ratio of different type tubules were recorded. It was revealed that, normally, granulocytomes were formed during utilization of necrotic tubules. Parasitic invasion of trematodes entailed swelling of the gland and a change in the proportion of tubules: the number of absorptive tubules decreased to 20%, the number of destructing and restoring tubules increased up to 60% and 12%, respectively.