Impact of Irgarol 1051 on the larval development and metamorphosis of Balanus amphitrite Darwin, the diatom Amphora coffeaformis and natural biofilm

The effect of Irgarol 1051 on the biofilm-forming diatom, Amphora coffeaformis, and on natural biofilm (NBF) was assessed. A reduction in the number of A. coffeaformis cells within a biofilm was observed after treatment with Irgarol 1051, confirming its role as an inhibitor of photosynthetic activity. The impact of this compound on the development of nauplii of Balanus amphitrite was evaluated through its impact on Chaetoceros calcitrans, which was provided as food for the larvae. A reduction in the number of cells of C. calcitrans was observed when treated with Irgarol 1051. When larvae of B. amphitrite were reared using C. calcitrans in the presence of Irgarol 1051, their mortality increased with an increase in the concentration of Irgarol 1051 (13% at 1 microg l(-1) to 40% at 1000 microg l(-1)) compared with the control (6%). Nauplii reared in the presence of Irgarol 1051 developed more slowly (6-7 days) compared with control larvae (4-5 days). Cyprid bioassay results indicated an increase in percentage metamorphosis (76%) when NBFs were treated with the highest concentration of Irgarol 1051, compared with untreated biofilm (28%). The enhanced rate of metamorphosis appeared to be related to an increase in bacterial numbers in the biofilm, which may have been due to lysis of diatoms caused by Irgarol 1051. A. coffeaformis biofilms grown in the presence of antibiotics showed a significant reduction in cell numbers, which on further treatment with Irgarol 1051 showed an increase in cell numbers. Thus, it can be hypothesised that A. coffeaformis cells that were subjected to stress twice may have expressed resistant genes. Furthermore, if plasmids are present in the biofilms, they may enhance transfer to the surviving cells making them more resistant to hostile conditions.     

Size of secretory granules from midgut cells of the stablefly, Stomoxys calcitrans

The size frequency distribution and number of secretory granules in the opaque zone midgut cells of the stablefly Stomoxys calcitrans have been determined using several independent methods. Average mean secretory granule diameter is 0.45 mum. Mean number of secretory granules per cell is 28,560. The diameter of the granules is compared to the size of secretory granules found in complementary vertebrate tissues and the consequences for the cell of the differences in the size of the granules is discussed.     

Winter feeding sites of hay in round bales as major developmental sites of Stomoxys calcitrans (Diptera: Muscidae) in pastures in spring and summer

The stable fly, Stomoxys calcitrans (L.), historically has been a pest of livestock in confined operations but seldom of animals on pastures or rangelands. In the past two decades, however, S. calcitrans has become a major pest of cattle and horses on pastures in the midwestern United States. Although there usually is an overabundance of diverse stable fly and house fly, Musca domestica L., larval habitats in confined livestock operations, no larval habitat for stable flies has been clearly identified in the pasture-range environment. Because the winter feeding of hay in round bales results in significant amounts of hay wastage that when mixed with manure, might develop into suitable larval habitats, this study evaluated these areas as developmental sites for the abundant stable flies in pastures. There was a trend for fly traps placed in the vicinity of hay feeding sites to catch more stable flies than those placed distant from these sites. Estimates of stable flies emerging from these sites ranged from 102 to 1225 flies per core sample (25 by 25 cm). The mean number of adult stable flies during May and June 2001 through 2004 correlated negatively with the average minimum temperatures during the preceding winter (November-February) but not with rainfall or temperatures during the spring. These results support the hypothesis that winter feeding sites of hay in round bales are the main source of stable flies in pastures.     

A new morphological character for the identification of Stomoxys calcitrans and S. niger (Diptera: Muscidae). Comparison of La Réunion island populations

On La Reunion Island (France), two morphological closely related species of stable flies (Diptera: Muscidae), living in the same environment, Stomoxys calcitrans and S. niger, are involved in the transmission of blood parasites to the livestock. To facilitate a rapid identification of both species in the field conditions, we highlighted a diagnostic morphological character not yet described: the length of the maxillary palpus. The study of three populations of S. calcitrans and two populations of S. niger, collected at various elevations, showed that the maxillary palpi of S. niger were significantly longer than in S. calcitrans, independent of sex. This character, easily visible in the field with a simple magnifying glass, has been confirmed on individuals of both species from West Africa.     

Epithelial innate immunity. A novel antimicrobial peptide with antiparasitic activity in the blood-sucking insect Stomoxys calcitrans

The gut epithelium is an essential interface in insects that transmit parasites. We investigated the role that local innate immunity might have on vector competence, taking Stomoxys calcitrans as a model. S. calcitrans is sympatric with tsetse flies, feeds on many of the same vertebrate hosts, and is thus regularly exposed to the trypanosomes that cause African sleeping sickness and nagana. Despite this, S. calcitrans is not a cyclical vector of these trypanosomes. Trypanosomes develop exclusively in the lumen of digestive organs, and so epithelial immune mechanisms, and in particular antimicrobial peptides (AMPs), may be the prime determinants of the fate of an infection. To investigate why S. calcitrans is not a cyclical vector of trypanosomes, we have looked in its midgut for AMPs with trypanolytic activity. We have identified a new AMP of 42 amino acids, which we named stomoxyn, constitutively expressed and secreted exclusively in the anterior midgut of S. calcitrans. It displays an amphipathic helical structure and exhibits a broad activity spectrum affecting the growth of microorganisms. Interestingly, this AMP exhibits trypanolytic activity to Trypanosoma brucei rhodesiense. We argue that stomoxyn may help to explain why S. calcitrans is not a vector of trypanosomes causing African sleeping sickness and nagana.     

Effect of autologous blood in the hypo-osmotic state on the immune system of the body

The analysis of the immune status of patients under neurodermatitis after autologous blood treatment has been performed. It has been found that positive therapeutic effect correlates with a number of immunologic parameters: the lysozyme content raises, the metabolic activity of neutrophilic cells increases, a number of T-lymphocytes grows to a certain degree, the balance of T-lymphocyte subpopulation normalizes.     

Sensory and behavioural responses of the stable fly Stomoxys calcitrans to rumen volatiles

Analysis of volatiles from rumen digesta by gas chromatography linked antennogram recordings from Stomoxys calcitrans (L) (Diptera: Muscidae) antennal receptor cells revealed about 30 electrophysiologically active constituents, the most important of which is dimethyl trisulphide with a sensory threshold in the femtogram range. The behavioural responses of S. calcitrans to five chemostimulants (dimethyl trisulphide, butanoic acid, p-cresol, oct-1-en-3-ol and skatole) were tested in a wind tunnel where activation and attraction of hungry flies to rumen volatiles were recorded. Dimethyl trisulphide, butanoic acid and p-cresol were found to attract S. calcitrans. This sensitivity to rumen volatile constituents, that also occur in animal wastes used for oviposition by Stomoxys spp., as well as in flowers used by stable flies as sources of nectar is discussed in the context of the behavioural ecology of these flies.     

On the selection for the modal chromosome number in Chinese hamster cells

Chinese hamster cell lines have a strong tendency to maintain as modal chromosome number the euploid one characteristic of the species. By following the evolution of karyotypes in cloned subpopulation it has been possible to show that these pseudodiploid constitutions are not simply established due to a selective advantage but they are actually generated in populations where pseudodiploid classes had been absent. The possible role of the mitotic apparatus in keeping and restoring the euploid number is discussed.     

Diversity of Stomoxys spp. (Diptera: Muscidae) and diurnal variations of activity of Stomoxys indicus and S. calcitrans in a farm, in Wang Nam Khiao District, Nakhon Ratchasima Province, Thailand

A study of species diversity of Stomoxys spp. and diurnal variations of activity of the most abundant was performed during a one year period at a local dairy cattle farm in Wang Nam Khiao District, Nakhon Ratchasima Province, Thailand. Four species of stomoxyine flies were morphologically identified, including Stomoxys indicus Picard 1908, S. calcitrans (Linnaeus 1758), S. sitiens Rondani 1873 and S. uruma Shinonaga and Kanao 1966. The most common species were S. indicus (50.2%) and S. calcitrans (49.5%). S. sitiens and S. uruma were found in small proportions (< 1%). The number of flies captured was significantly different among the three seasons with the greatest number in the rainy season (mean = 66%; df = 2, P < 0.05). The variations of diurnal activity were observed during different period of times (06:00 to 18:00) during three seasons. Both sexes of S. indicus and males of S. calcitrans showed unimodal activity pattern in cool and summer seasons. But a bimodal activity pattern was recorded in rainy season. For females S. calcitrans, a unimodal peak of activity was observed in cool season and a constant variation of activity all along the day in summer and rainy seasons, with an increase from the morning to the evening. A better understanding of stomoxyine fly behavior, especially the daily flight activity, can assist in prioritization and design of appropriate vector prevention and control strategies.     

Chemostimuli implicated in selection of oviposition substrates by the stable fly Stomoxys calcitrans

Horse and cow dung were tested as substrates for oviposition by the stable fly Stomoxys calcitrans (L) (Diptera: Muscidae) in laboratory cages. Odour alone from either horse or cow dung was sufficient to attract flies for oviposition. This was confirmed in wind tunnel experiments, where both horse and cow dung were shown to attract gravid stable flies. However, when S. calcitrans was offered a choice between these two oviposition substrates, flies always chose horse dung over cow dung, both when allowed to contact the substrates and when relying on dung odour alone. Analyses of volatile compounds emanating from horse and cow dung by gas chromatography linked antennogram recordings from S. calcitrans antennae revealed no differences in the chemostimuli released from the two substrates. The predominant chemostimulant compounds in both substrates were carboxylic acids (butanoic acid), alcohols (oct-1-en-3-ol), aldehydes (decanal), ketones (octan-3-one), phenols (p-cresol), indoles (skatole), terpenes (beta-caryophyllene) and sulphides (dimethyl trisulphide). Higher levels (20-40 p.p.m.) of carbon dioxide were recorded over horse dung compared with cow dung, a factor that may contribute to the preference exhibited by S. calcitrans for this substrate for oviposition.     

A note on the effect of controlling stable flies (Stomoxys calcitrans) in the resting activity and pen distribution of dairy cows

This study investigated the effect of controlling stable flies (Stomoxys calcitrans) on the number of dairy cows lying down and their pen distribution. The study randomly assigned 80 Holstein cows to 1 of 2 groups. The treated group (T) included cows individually sprayed with insecticide when found with an average of 10 stable flies per cow; in the control group (C), cows received no application of insecticide. The pen had 4 equal-size areas: (a) feeding, (b) drinking and sunny, (c) covered, and (d) manure. The study recorded the number of cows lying and the area of the pen where the nonhuman animal was located. The study found no difference (p > .05) between the proportion of T and C cows lying. However, cows preferred to lie down in pen area 3 when fewer than 10 stable flies per cow were found. Area 4 was the most avoided section of the pen, except when a high incidence of flies was present. The study concluded that high populations of S. calcitrans (> 10 flies per cow) did not affect the number of dairy cows lying down. However, it modified site preferences for lying.     

Enumeration of T lymphocyte subsets by monoclonal antibodies in young and aged humans

The percentage and absolute number of peripheral blood mononuclear cells reactive with monoclonal antibodies identifying mature thymocytes and T cells (T3+), helper/inducer cells (T4+), and suppressor/cytotoxic cells (T8+) was determined in 19 young (mean age 35 yr) and 31 elderly (mean age 72 yr) individuals. The percent representation but not the absolute number of T cells (T3+) declined significantly (p less than 0.001) in the elderly, and the decline was attributable to both an absolute and relative decrease in the representation of the subpopulation of cytotoxic/suppressor (T8+) cells. The percentage and number of helper/inducer (T4+) T cells was comparable in both age groups.     

A Note on the Effect of Controlling Stable Flies (Stomoxys Calcitrans) in the Resting Activity and Pen Distribution of Dairy Cows

This study investigated the effect of controlling stable flies (Stomoxys calcitrans) on the number of dairy cows lying down and their pen distribution. The study randomly assigned 80 Holstein cows to 1 of 2 groups. The treated group (T) included cows individually sprayed with insecticide when found with an average of 10 stable flies per cow; in the control group (C), cows received no application of insecticide. The pen had 4 equal-size areas: (a) feeding, (b) drinking and sunny, (c) covered, and (d) manure. The study recorded the number of cows lying and the area of the pen where the nonhuman animal was located. The study found no difference (p > .05) between the proportion of T and C cows lying. However, cows preferred to lie down in pen area 3 when fewer than 10 stable flies per cow were found. Area 4 was the most avoided section of the pen, except when a high incidence of flies was present. The study concluded that high populations of S. calcitrans (> 10 flies per cow) did not affect the number of dairy cows lying down. However, it modified site preferences for lying.     

Identification of the intermediate hosts of Habronema microstoma and Habronema muscae under field conditions

Abstract.  A polymerase chain reaction (PCR)-based assay was used for the specific detection of Habronema microstoma and Habronema muscae (Nematoda, Spirurida) in order to identify the intermediate hosts of both nematode species under field conditions. A total of 1087 netted and 165 laboratory-bred flies were tested. Flies were identified as Musca domestica Linnaeus 1758, Musca autumnalis De Geer 1776, Haematobia irritans (Linnaeus 1758), Haematobia titillans (De Geer 1907) and Stomoxys calcitrans (Linnaeus 1758) (Muscidae). Genomic DNA was extracted from pools of fly heads, thoraces and abdomens, and 703 samples were subjected to a duplex two-step semi-nested PCR assay to specifically detect diagnostic regions within the ribosomal ITS2 sequence of both H. microstoma and H. muscae . Stomoxys calcitrans specimens were positive for H. microstoma DNA and M. domestica specimens were positive for H. muscae DNA. In particular, PCR-positive samples derived from both farm-netted and laboratory-bred flies. The present study represents the first evidence of the vectorial competence of different fly species as intermediate hosts of Habronema stomachworms under field conditions. We discuss the roles of S. calcitrans and M. domestica in transmitting H. microstoma and H. muscae .     

Quantitative evidence for merocrine secretion in insect midgut cells

The occurrence of apical extrusions from cells has been observed on a large number of occasions in a wide variety of histological investigations. But whether the extrusions observed are a normal physiological event or are artefacts of the experimental procedures adopted has been the subject of much controversy. Lehane (1976a) described the extrusion of apical portions of the opaque zone midgut cells of Stomoxys calcitrans which contained large numbers of storage granules (merocrine secretion). This process involved the cell in the loss of cytoplasm, mitochondria and membranes as well as secretory products. If merocrine secretion is a normal physiological event rather than a fixation artefact then large falls should occur in the quantities of these components in the cell following the secretion burst. Using stereological methods the opaque zone cells were quantitatively described before and after the period in which merocrine secretion occurs (using these timings any possibility of producing apical extrusions as fixation artefacts was avoided). The results are entirely consistent with merocrine secretion having occurred. The volume of the cytoplasm falls by 23.74%, mitochondria by 30.55% and the total area of membrane in the cell by 32.59%.     

Retention and attempted mechanical transmission of Ehrlichia risticii by Stomoxys calcitrans

Abstract. The ability of adult Stomoxys calcitrans (L.) (Diptera: Muscidae) to retain viable Ehrlichia risticii (Rickettsiaceae), the aetiologic agent of Potomac horse fever (PHF), and mechanically transmit the pathogen from citrated bovine blood artificially infected with E.risticii to susceptible mice was studied. Viable E. risticii were found in the digestive tract of S.calcitrans 3h after the flies had engorged to repletion on infected blood; however, no E. risticii were detected in flies ≥2 days after feeding. Subsequently, groups of S.calcitrans were fed for 20 s on infected blood, then fed to repletion on mice 30, 60,130, 180 or 220 min after having feeding interrupted. Mice displayed no clinical signs of PHF and did not produce anti- E. risticii antibodies when assayed 30 days after S. calcitrans had fed. Although S.calcitrans were able to harbour viable E.risticii for at least 3h, transmission of the disease agent to susceptible mice during interrupted feeding was not demonstrated under these experimental conditions.     

Effect of host behaviour on host preference in Stomoxys calcitrans

Field observations suggest that, in the U.K., cattle are the preferred host of Stomoxys calcitrans (L.), followed by horses. Differences were observed in the numbers of flies feeding on individual animals both in the field and under controlled conditions. Analysis of the behaviour of four Friesian calves under attack from S. calcitrans in controlled conditions revealed that the differences in the levels of attack between individual hosts are dependent on the reactions of the host when under attack. Those hosts which respond vigorously by tail flicks, foot stamps and head-swings suffer less from attack by S. calcitrans than their more placid contemporaries.     

Twenty-three new microsatellite loci in the stable fly, Stomoxys calcitrans (L.) (Diptera: Muscidae)

The stable fly, Stomoxys calcitrans (L.), is a significant pest of cattle. Twenty-three microsatellite markers were isolated from a repeat-enriched genomic library of S. calcitrans. We characterized variation at these markers and found that 17 loci were polymorphic in two fly populations from Florida. Two to nine alleles were observed among the variable microsatellite loci and expected heterozygosities ranged from 0.03704 to 0.85115. These markers will be useful for characterizing population genetic differentiation and for tracking the migration patterns of stable flies in the USA and worldwide.