Effects of Plant Contact, Inoculation Pattern, Leaf Wetness Regime, and Nitrogen Supply on Inoculum Efficiency in Rice Sheath Blight

An experimental design was developed which allowed the measurement of inoculum efficiency (IE) represented by lesions of a leaf-borne disease such as rice sheath blight. In this design. IE is measured as the ratio of newly established lesions on trap plants relative to the inoculum present in a canopy, i.e. lesions artificially established on source plants, IE of the rice sheath blight pathogen, Rhizoctonia solani , was studied under semi-controlled conditions in which the effects of the following factors were measured; contact frequency among host tissues, amount and location of inoculum in the canopy, leaf wetness regime, and nitrogen content of the host plant. The conduciveness of these factors to sheath blight infection was measured in terms of IE which was expressed as the ratio of the density of daughter lesions on trap rice hills to the density of mother lesions on the inoculated quadrat hills. IE generally declined with the three successive batches of trap hills used in the experiments. It was significantly higher at closer plant spacings and under interrupted leaf wetness regimes. IE was not affected by the amount, nor by the location, of inoculum (mother lesions) in the canopy, but was significantly lower in hills with high total nitrogen content.     

Brugia pahangi and Dirofilaria immitis: experimental infections in the ferret, Mustela putorius furo.

Ferrets were inoculated with 160 third-stage larvae of the filarial nematode Brugia pahangi, followed 23 days later by 15 larvae of another filarial nematode, Dirofilaria immitis. Other ferrets received only one of these species. Microfilaremia developed in some ferrets with single infections of each species and in some ferrets with dual infections. The nature of the experiment did not permit a thorough study of microfilaremia, but B. pahangi microfilariae were found in numbers as high as 15,650/ml. At necropsy, approximately 8 months after inoculation, adult B. pahangi were recovered from the lymphatic vessels of all 8 ferrets inoculated only with that species, the recovery rate (based on 6 animals only) varying from 2 to 50% of the inoculum (mean 25%). Adult D. immitis were recovered from the heart of all three ferrets inoculated only with that species, the recovery rate being 7, 47, and 60% (mean 38%) of the inoculum. All 5 ferrets inoculated with both species yielded both adult B. pahangi (6 to 23%, mean 16% of inoculum) and adult D. immitis (13 to 67%, mean 37% of inoculum). It is concluded that the ferret is highly susceptible to both species and that concurrent infections with both species may readily be established.     

Effect of leaf age and inoculum concentration on the symptoms produced by Gymnosporangiumjuniperi-virginianae on apple

Gymnosporangium juniperi-virginianae, the incitant of cedar apple rust, induced the greatest number of lesions (flecks, pycnial lesions, aecial lesions) on 6- and 8-day-old leaves of single-shoot potted apple plants inoculated and incubated under controlled conditions. The largest pycnial lesions developed on 2- and 4-day-old leaves and the largest aecial lesions on 6-day-old leaves. The number of lesions was positively correlated with the concentration of basidiospores in the inoculum, but lesion size was constant at different inoculum levels. Elevated inoculum concentrations failed to induce pycnia on cv. McIntosh which normally bears only flecks; very low concentrations induced the development of pycnia but not aecia on cv. Rome Beauty, which normally bears aecia. An ‘infection rating’, log₁₀ (10 nd²), where n is the highest number of pycnial lesions per leaf and d is the largest mean diameter of pycnial lesions on any leaf, calculated for four cultivars in greenhouse tests showed a positive correlation with a ‘susceptibility rating’ developed from observations of natural infection of these cultivars in the field.     

Incorporation of radioactive precursors into filarial larvae of Brugia developing in susceptible and refractory mosquitoes.

The incorporation of tritiated precursors injected into mosquito hosts parasitized by developing filarial larvae of Brugia patei has been studied by autoradiography in 2 species of mosquito, Aedes togoi in which filarial development was normal and Anopheles labranchiae atroparvus in which filarial development was abnormal. In both mosquito hosts there was significant incorporation into 4--5-day-old developing larvae of uridine and amino acids (isoleucine, leucine, valine, arginine, lysine, cystine, methionine, phenylalanine, tyrosine, tryptophan, histidine, and proline), although lower incorporation of methionine, tyrosine, and tryptophan was found during abnormal development. No incorporation of thymidine, hydroxytryptophan, dopa, or carbohydrate was found at this stage of larval development. Some incorporation of glucose and dopa was found in or around earlier stages of development in An. l. atroparvus. Mosquito flight muscle showed lower incorporation of glucose, but not of amino acids, around the site of filarial parasite development. The flight muscle of An. l. atroparvus showed a higher level of incorporation of lysine compared to that in A. togoi and higher levels of lysine and valine were found in the abnormally developing filarial larvae in the refractory mosquito.     

Syphacia obvelata in the Mongolian gerbil (Meriones unguiculatus): natural occurrence and experimental transmission

An adult from a research colony and a litter of 5-week-old Mongolian gerbils (Meriones unguiculatus) from a pet store were found to have pinworms identified as Syphacia obvelata. The infected gerbils were allowed to cohabitate with uninfected gerbils. Similarly, infected gerbils were caged with uninfected mice and infected mice with uninfected gerbils. Results of these studies showed that Syphacia obvelata can be transmitted from gerbil to gerbil, gerbil to mouse, and mouse to gerbil.     

In vitro chemotaxis of Brugia pahangi infective larvae to the sera and hemolymph of mammals and lower animals

The jird (Mongolian gerbil) is a highly susceptible experimental host for the lymphatic filarial nematode, Brugia pahangi. The chemotactic activity of serum from this host for B. pahangi infective larvae was compared in vitro to that of sera or hemolymph of a wide variety of other organisms including mammals, reptiles, fishes and invertebrates. The range of the Chemotactic Index (CI) was from 96.0 for the jird to 56.2 for a snail. An average of CI of saline control was 4.5. Significant chemotactic activity was present in many organisms, especially mammals, but was not closely related to either the phylogenetic position of the organism and to its known susceptibility as definitive host for B. pahangi. Migratory response was diminished in a consistent way by serial dilution of sera of humans, jirds and fetal bovine serum. Pre-incubation of larvae in fetal bovine serum inhibited migration, especially towards the sera of humans. Inhibition could be reversed by rinsing larvae in saline, longer rinse periods resulting in greater recovery of CI. These results are the first to suggest the activity of the specific amphid chemoreceptors in the chemotaxis of the infective larvae of B. pahangi.     

Association of ABO and Rh(D) blood groups with filariasis

ABO and Rh blood groups in 344 filarial patients and 320 controls matched with respect to age, community and residence are reported. An excess of B and a deficiency of AB was observed among the filarial patients. The relative risks for the B and AB were 1.53 and 0.36, respectively. Only males showed clearly significant risks. The Rh(D) blood groups revealed no association with filariasis.     

The effect of Brugia pahangi infection on survival of susceptible and refractory species of the Aedes scutellaris complex

Life table statistics were used to examine the survival functions of filarial susceptible and refractory species of the Aedes scutellaris (Walker) group of mosquitoes, following infection with high and moderate doses of Brugia pahangi (Buckley & Edeson). Survivorship curves and hazard function curves were generated, and the median survival times and the proportions of mosquitoes surviving beyond the extrinsic incubation period of the parasite were determined. In the susceptible populations of Aedes polynesiensis Marks, Ae. pseudoscutellaris (Theobald) and Ae.tabu Ramalingam & Belkin a dose-response relationship was detected between parasite load and mortality. This relationship was characterized by a significant reduction in the proportions of infected female mosquitoes surviving at days 1 and 9 postinfection, reduction in the median survival times and an increase in the hazard rates as the infectious dose increased. The survival of the refractory species, Ae.alcasidi Huang and Ae.katherinensis Woodhill was not significantly affected by the infection. A positive correlation between microfilaraemia in the vertebrate host and parasite load in the susceptible mosquito populations was also observed. Regression analysis of the number of parasites recovered from susceptible mosquitoes at the time of death showed that mosquitoes at highest risk of dying harboured from 11.6 to 19.4 infective larvae when fed on a gerbil with sixty-five microfilariae per 20 microliters blood; this resulted in 34.4-40.2% mortality by day 9 postinfection. A mean number of 32.6-46.9 infective larvae was observed when these populations were exposed to a gerbil with a microfilaraemia of 150 mf/20 microliters and resulted in 72.8% to 80% mortality in these populations. Viable infective larvae were recovered from infected mosquitoes up to 50 days postinfection.     

The infection of potatoes by Phoma exigua

The pathogenicities of Phoma exigua var. foveata and P. exigua var. exigua were compared by determining the relationship between incidence of infection of whole tubers or tuber pieces and various inoculum levels in soil. In natural soil var. foveata was more pathogenic than var. exigua, but in sterilized soil there were no clear differences between the two. Both varieties were isolated from some lesions derived from inoculum to which only one variety had been added indicating either natural contamination of soil or latent infection in tubers; the proportion of such lesions increased at the lower inoculum concentrations. In studies where potato plants were grown in infested soils, the incidence of stem disease and tuber disease increased at increased inoculum concentrations of var. foveata; var. exigua was not added to the inoculum but was isolated from some lesions, the proportions of such lesions again increasing at lower inoculum concentrations. The infectivity of the infested soil, assessed by a tuber-baiting method, declined soon after planting but increased prior to harvest. Laboratory studies indicated that in natural soil the infectivity was highest after 2–3 months and lowest after 10 months incubation, but in sterilized soil infectivity was highest after 5 months and lowest after 7 months incubation.     

Quantitative appraisal of murine filariasis confirms host strain differences but reveals that BALB/c females are more susceptible than males to Litomosoides sigmodontis

Litomosoides sigmodontis, a rodent filarial nematode, can infect inbred laboratory mice, with full development to patency in the BALB/c strain. Strains such as C57BL/6 are considered resistant, because although filarial development can occur, circulating microfilariae are never detected. This model system has, for the first time, allowed the power of murine immunology to be applied to fundamental questions regarding susceptibility to filarial nematode infection. As this is a relatively new model, many aspects of the biology remain to be discovered or more clearly defined. We undertook a major analysis of 85 experiments, to quantitatively assess differences in filarial survival and reproduction in male versus female and BALB/c versus C57BL/6 mice over the full course of infection. This large dataset provided hard statistical support for previous qualitative reviews, including observations that the resistant phenotype of C57BL/6 mice is detectable as early as 10 days postinfection (dpi). An unexpected finding, however, was that filarial survival was reduced in male BALB/c mice compared to their female counterparts. Worm recovery as well as the prevalence and density of microfilariae were higher in female compared with male BALB/c mice. Therefore, L. sigmodontis bucks the filarial trend of increased susceptibility in males. This could be partially explained by the different anatomical locations of adult L. sigmodontis versus lymphatic filariae. Interestingly, the effects of BALB/c sex upon microfilaremia were independent of worm number. In summary, this study has significantly refined our understanding of the host-L. sigmodontis relationship and, critically, has challenged the dogma that males are more susceptible to filarial infection.     

A comparative analysis of the properties of bacteria in the genus Salmonella isolated from children in Latvia]

Since social and economic changes in Latvia in 1991 the incidence of salmonellosis dramatically increased: more than 500 pediatric cases are registered every year. Specification of the properties of the Salmonella strains isolated in Latvia was undertaken. The study demonstrated that acute salmonellosis in pediatric inpatients was mainly due to S.typhimurium (78 per cent) and only in 22 per cent of the patients it was due to S.enteritidis. All the S.typhimurium isolates showed high antibiotic resistance defined by specific extended spectrum beta-lactamase (CTX-M-5).     

Differential susceptibility to Brugia pahangi infection in Mongolian gerbils (Meriones unguiculatus) of different coat colour

The influence of intraspecific host variables on the response to parasitic infections is an important aspect of host-parasite relationships, yet little is known about this aspect of filariasis for lack of a model. This study presents coat colour mutants of the Mongolian gerbil (Meriones unguiculatus) as potential new models for research into the effects of host genetic variation on response to filarial infection. Peak level of microfilaraemia, eosinophil response, body weight and degree of splenomegaly in gerbils infected with Brugia pahangi varied with agouti, albino, and black coat colour. These results suggested that coat colour-related genes might influence host immune response to developmental stages of the parasite and eosinophil-mediated reaction might cause host damage.     

Pathology of mesenteric lymph nodes of buffaloes and cattle with special reference to salmonellosis

Enlarged mesenteric lymph nodes were collected from ninety buffaloes and sixty cattle slaughtered at Faisalabad abattoir. Among these, salmonellae were isolated from lymph nodes of 32 (21.33%) animals. Maximum preponderance of salmonellosis was recorded in animals over two years of age. Enlargement, pale to dark red in color, increased consistency and even calcification were the main gross pathological lesions. Histopathological lesions included thickened capsule, typical lymphofollicular reaction, accumulation of oedematous fluid and haemorrhages.     

Preliminary results on the effect of tetracycline on the embryogenesis and symbiotic bacteria (Wolbachia) of Dirofilaria immitis. An update and discussion.

The distribution and phylogeny of Wolbachia in filarial species suggests that these endosymbiotic bacteria may be important in the biology of their filarial hosts. An experiment to falsify this hypothesis would be to treat filarial worms with antibiotics which are active against intracellular bacteria. Indeed, it has already been shown that tetracycline treatment inhibits development in a model filarial species (Brugia pahangi) at different stages of the life cycle, in both mosquito and mammalian hosts. Here we discuss these previous data and present new results on the effect of tetracycline on the embryogenesis of the canine filaria Dirofilaria immitis.     

Effect of immunostimulatory oligodeoxynucleotides on host responses and the establishment of Brugia pahangi in Mongolian gerbils (Meriones unguiculatus)

Infection of humans with filarial parasites has long been associated with the maintenance of a dominant Th2-type host immune response. This is reflected by increases in interleukin (IL)-4- and IL-5-producing T cells, elevated immunoglobulin (Ig)E and IgG4 levels, and a pronounced eosinophilia. The Mongolian gerbil (Meriones unguiculatus) is permissive for the filarial nematodes Brugia malayi and B. pahangi. As in humans, persistent microfilaremic infections of gerbils with Brugia spp. results in increases in Th2 cytokines such as IL-4 and IL-5. The association of dominant Th2 cytokine profiles with the maintenance of infection suggests that the introduction of Brugia spp. into a strongly Th1-biased environment may adversely affect parasite establishment. Indeed, studies conducted in mice with B. malayi suggest that depleting Th1 effectors such as interferon (IFN)-gamma and nitric oxide results in increased worm recoveries. In the present studies, the Mongolian gerbil was used as a model to investigate the effect of a dominant Th1 cytokine environment on the establishment of B. pahangi. Intraperitoneal (i.p.) administration of immunostimulatory oligodeoxynucleotide (IS ODN) induced the production of IFN-gamma in the peritoneal exudate cells and spleen of gerbils. The presence of IFN-gamma at the time of B. pahangi infection did result in an altered host immune response to B. pahangi. Gerbils that received IS ODN before i.p. B. pahangi infections showed lower levels of the Th2-type cytokines IL-4 and IL-5, compared with animals that received B. pahangi alone (0 + Bp). This alteration in cytokine profile, however, did not alter the establishment or development of B. pahangi in the peritoneal cavity. Furthermore, there was no difference in the granulomatous response of gerbils to soluble adult B. pahangi antigen bound to beads embolized in their lungs, regardless of treatment group, suggesting that IL-4 and IL-5 are not essential contributors to the systemic host inflammatory response to B. pahangi in this model.     

Brugia pahangi: in vivo tissue migration of early L3 alters gene expression

Events occurring during early filarial nematode migrations are central to parasite establishment but rarely studied. Brugia pahangi larvae injected intradermal (ID) into the hind limb of the gerbil (Meriones unguiculatus) can be recovered from the popliteal lymph node (POP) at 3 days post-infection (DPI). They have been designated migrating larvae (IDL3). Alternatively, L3 recovered at 3DPI from the peritoneal cavity (IPL3) do not migrate. Subtracted cDNA libraries using IDL3 and IPL3 revealed distinct gene profiles between IDL3 and IPL3. Troponin-c was significantly upregulated in IDL3, while Cathepsin L was significantly increased in IPL3. Differences in mRNA levels were also observed with these and other genes between IDL3, IPL3 and L3 isolated from mosquitoes (VL3). These data suggest that migratory activity, exposure to potentially different host environments and/or host location may be important external factors in influencing larval gene expression.     

Inhibition of spermatogenesis in gerbil (Meriones hurrianae) hedgehog (Hemiechinus auritus collaris) and mice after methallibure (ICI 33828) treatment

The effect of methallibure (ICI 33828) on spermatogenesis was studied in the gerbil, hedgehog, and mouse. Injection of ICI 33828, at a dose of 100 mg/kg for 10-25 days, caused testicular lesions in the gerbil and hedgehog, and spermatogonia were absent in the seminiferous e pithelium. Distinct hypertrophy of the Leydig cells was evident. Acces sory sex gland activity was suppressed. It is possible that ICI 33828 impairs testicular androgen production and luteinizing hormone (LH) activity, as suggested by reduced seminal veisicle weight and reduced concentrations of DNA and RNA. Levels of follicle stimulating hormone and LH were considerably reduced. Both adult male gerbils and hedgehogs were rendered sterile by ICI 33828 at the dose studied, while mice were relatively unaffected.     

The mosquito fauna of Smolensk and Kaluga provinces

The mosquitos collected in Smolensk and Kaluga provinces in 1985 by the expedition team of the D. I. Ivanovski? Institute of Virology were identified by V. N. Danilov in 1986. Among the females of mosquitos collected in Smolensk Province four genera and 16 species were recorded; two genera (Culiseta, Mansonia) and 9 species were new ones for this region. Taking in account the reference data (excluding Anopheles hircanus and Aedes behningi, the presence of which here is doubtful) there 22 species of mosquitos of five genera (Anopheles--3, Aedes--16, Mansonia, Culiseta, Culex--one of each) are recently known in Smolensk Province. Among the females of mosquitos collected in Kaluga Province, two genera and 8 species were found including one genus (Mansonia) and three species being a new ones for this region. Together with the reference data, there 17 species of mosquitos of four genera (Anopheles--2, Aedes--13, Mansonia, Culex--one of each) are known now for Kaluga Province.     

Components of resistance to Cercospora arachidicola and Cercosporidium personatum in groundnuts*

The reactions of four groundnut cultivars to inoculation with two leaf spot pathogens (Cercospora arachidicola and Cercospodirium personatum) were investigated. Resistant cultivars showed longer latent periods, reduced sporulation and less defoliation; however, none of the cultivars was resistant to both pathogens. Numbers of lesions, time to leaflet loss and the percentage of spores which produced lesions were influenced by the concentration of C. personatum inoculum applied to the leaves and there were significant cultivar differences in lesion numbers at the lowest inoculum level. Only the first two characters were affected by applied spore concentration in the study with C. arachidicola. Generally it was found that lower inoculum levels gave a better separation of cultivars. The level of resistance was similar to that found in the slow resting reaction of cereal crops and should be useful in a groundnut breeding programme.     

The pathways of energy generation in filarial parasites

It has been commonly accepted that most adult filarial parasites use the glycolytic breakdown of carbohydrates to lactate as a preferred route to supply their energy requirements. Their ability to catabolize glucose by oxygen-dependent pathways is rather limited. An exception to this is the rodent filarial species Litomosoides carinii, which requires a unique type of aerobic glucose metabolism to maintain motor activity and survival. However, the prominent role of carbohydrates as energy substrates for filariids may no longer be tenable. Recent studies have shown that glutamine is a major energy source in filarial worms and that a fully oxidative mitochondrial metabolism can be employed for the utilization of this substrate.