Regression of the lateral oviducts during the larval-adult transformation of the reproductive system of Melipona quadrifasciata and Frieseomelitta varia

Toward the end of the larval phase (pre-pupa), the reproductive systems of Melipona quadrifasciata and Frieseomelitta varia workers are anatomically similar. Scanning electron microscopy showed that during this developmental phase the right and left ovaries are fused and form a heart-shaped structure located above the midgut. Each ovary is connected to the genital chamber by a long and slender lateral oviduct. During pupal development, the lateral oviducts of workers from both species become extremely reduced due to a drastic process of cell death, as shown by transmission electron microscopy. During the lateral oviduct shortening, their simple columnar epithelial cells show some signs of apoptosis in addition to necrosis. Cell death was characterized by cytoplasmic vesiculation, peculiar accumulation of glycogen, and dilation of cytoplasmic organelles such as mitochondria and rough endoplasmic reticulum. The nuclei, at first irregularly contoured, became swollen, with chromatin flocculation and various areas of condensed chromatin next to the nuclear envelope. At the end of the pupal phase, deep recesses marked the nuclei. At emergence, worker and queen reproductive systems showed marked differences, although reduction in the lateral oviducts was an event occurring in both castes. However, in queens the ovarioles increased in length and the spermatheca was larger than that of workers. At the external anatomical level, the reproductive system of workers and queens could be distinguished in the white- and pink-eyed pupal phase. The metamorphic function of the death of lateral oviduct cells, with consequent oviduct shortening, is discussed in terms of the anatomical reorganization of the reproductive system and of the ventrolateral positioning of adult worker bee ovaries. (C) 2000 Wiley-Liss, Inc.

Skin test and tick immune status in susceptible and resistant cattle in Brazil

A cutaneous hypersensitivity test (CHT) was used to correlate host resistance to ticks and type of reaction elicited to unfed larval extract-ULE of the cattle tick Boophilus microplus in European and Indian cattle. Twenty calves were separated into four groups of five animals each: naïve or preinfested Indian or European cattle. CHT was induced by intradermal inoculation of 0.1 ml of ULE cattle tick B. microplus (50 μg protein) in the calf ear. Ear thickness was measured using calipers before and 10 min, 1, 2, 6, 18, 24, 48, 72, 96, and 144 h postinoculation (PI). Preinfested European calves showed only an immediate type reaction with maximum response (75% increase in ear thickness) at 10 min PI. On the other hand, preinfested Indian calves presented an immediate response with maximum reaction (70% increase in ear thickness) between 10 min and one hour PI, and a delayed type reaction at 72 h PI (60% increase in ear thickness). These results point out the crucial role of the cellular immune response of cattle in the expression of resistance to cattle tick B. microplus. Skin test might be useful in the ranking of cattle according to the susceptibility/resistance to ticks.

Susceptibility of tangerines to citrus variegated chlorosis

Citrus variegated chlorosis (CVC), a citrus disease first discovered in Brazil in 1987, is caused by the bacterium Xylella fastidiosa and transmitted by sharpshooters and budwood. Since the disease affects almost all sweet orange cultivars, it has become one of the most serious problems for Brazilian citriculture. To evaluate their resistance to CVC disease, fifteen tangerines or mandarins (C. reticulata Blanco) and their hybrids were grafted on Rangpur lime (C. limonia Osb.) and inoculated with CVC-contaminated Pera sweet orange (C. sinensis (L.) Osb.) by twig grafting in a greenhouse. Tangerines and their hybrids Wilking, Fortune, Sunki, Ellendale, Orlando tangelo, Nunes clementine, Nova, Sun Shu Sha Kat, Suenkat, and Batangas showed CVC leaf symptoms and gave positive results on enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) (with specific primers for X. fastidiosa), indicating that they are susceptible to CVC. Although X. fastidiosa bacteria were detected by ELISA and PCR in inoculated plants of tangerines Cravo and Oneco, no CVC leaf symptoms were observed on these two cultivars, suggesting that they are tolerant to the disease. CVC leaf symptoms were not observed and X. fastidiosa was not detected in tangerine Dancy and mandarins Okitsu satsuma and Ponkan after inoculation, showing that they are resistant to the disease.

Larval development of Epialtus bituperculatus H. Milne Edwards, 1834 (Crustacea: Decapoda: Brachyura: Majidae) with comments on majid larvae from the southwestern Atlantic

The larval development of the spider crab Epialtus bituberculatus H. Milne Edwards which lives on rocky shores with algae such as Sargassum and Hypeneia, is described. Larvae were obtained from ovigerous females collected in Ubatuba, State of São Paulo, Brazil. Rearing was carried out at 24 ± 1°C, with an average salinity of 35%. Larval development consists of two zoeal stages and one megalopa. Zoeal development was completed in 9.5 days. Analysis indicated that zoeae of E. bituberculatus are very similar to those of E. brasiliensis Dana and Acanthonyx scutiformis (Dana). Differences noted between these species pertain to the setation of the carapace, maxillule and second maxilliped. The main morphological features useful for identification are presented together with a summary of features that characterize larvae of majid subfamilies in Brazil. A key for the identification of southwestern Atlantic majid zoeae to the family level is provided.

Oviposition behavior of Ceratitis capitata wiedemann (Diptera: Tephritidae): Association between oviposition preference and larval performance in individual females

The oviposition preference and larval performance of females of Ceratitis capitata (Wiedemann) were examined. Wild or laboratory-reared females were tested for oviposition preference in relation to papaya fruit in different stages of ripeness, as well as to two different fruits: papaya and apple. The results demonstrated that both laboratory-reared and wild females preferred ripe papaya fruit compared to unripe fruit for oviposition, and that ripe papaya was the best for larval performance. When oviposition preference was tested with different fruits (papaya and apple), both wild and laboratory-reared larvae developed better in papaya. However, only wild females showed a preference for oviposition on papaya as opposed to apple. Laboratory-reared females showed no oviposition preference on fruits that resulted in better larval performance.

Number of larval instars of the ant Pachycondyla (=Neoponera) villosa (Formicidae: Ponerinae) determined by the rule of Dyar

In this work we present the number of larval instars in the Ponerinae ant Pachycondyla (=Neoponera) villosa. The analysis of maximal head capsule width measurement of 147 larvae was made. Four larval instars were measured: 1st instar the cephalic capsule varied from 0.18mm to 0.22mm; 2nd instar from 0.23mm to 0.27mm; 3rd instar from 0.30mm to 0.33mm and the 4th instar varied from 0.35mm to 0.38mm. The mean growth rate was 1.2375 according to the rule of Dyar. We also reviewed the number of larval instars for 35 ant species.

Chemical analysis of different larval instars and tissues of Pachycondyla (=Neoponera) villosa (Hymenoptera: Formicidae, Ponerinae)

Chemical analyses of complete larvae of the first to third instar and cuticle, fat body and salivary glands extracts of fourth instar larvae using gas chromatography and gas chromatography-mass spectrometry, were performed upon Pachycondyla villosa. The results revealed that P. villosa larvae do not produce a pheromone, as only fatty acids and n-alkanes were detected. After quantifying the identified compounds, it was determined that the fat body is the main place of storage and/or production of the cuticular hydrocarbons. It was also observed that the absolute quantity of cuticular hydrocarbons increases progressively during larval development. Inferences about the transport behavior of matured larvae to the pupation place and the colony odor are discussed.

Ultrastructure of the salivary glands of Pachycondyla (=Neoponera) villosa (Fabricius) (Formicidae: Ponerinae): Functional changes during the last larval instar

The cells of secretory region of the salivary glands of Pachycondyla (=Neoponera) villosa at the time of enzyme production presents the basal cellular membranes profusely folded and the intercellular junctional membranes present a few enlarged spaces. The rough endoplasmic reticulum and the Golgi bodies shift from being flat and small vesicular cisternae to enlarged vesicular cisternae according to the cell physiological state and characterize an asynchronic cell cycle. Enzymes are released into the lumen by microapocrine secretion. The stage of silk production is detected after a behavioral act, when the nurse worker separates the mature larva. At this time, the salivary gland cells present only one physiological state (synchronized secretory cycle): this state was characterized by basal cellular membrane poorly folded, intercellular junctions presenting some small spaces, rough endoplasmic reticulum compounded by flat cistenae, enlarged Golgi bodies with fibrous material inside and a few secretory vesicles containing silk, which undergo exocytosis. The silk in the lumen shows 2 forms: tactoid and flocculent material.

Ovarian growth during larval development of queen and worker of Apis mellifera (Hymenoptera: Apidae): a morphometric and histological study.

The present work reports the differences between the ovarian grow in queen and worker larvae of A. mellifera, from the start of differential feeding. The observations made of the growth rates in larvae of both castes showed that the queen and worker larvae have the same rates of cephalic capsule growth from one instar to another but the weight gain is greater in queens. In the same way, the draw areas of ovaries of queens increase more and continuously, while from the 5th instar on the ovaries of workers decrease in size. The decrease is due to a loss of ovariole numbers that starts early in the worker larvae and increases in the 4th-5th instar. The ovarian shape in queens and workers became different in the last larval instars.

Effects of nitrate concentration on larval development of the giant river prawn, Macrobrachium rosenbergii

The effect of nitrate concentration on giant river prawn, Macrobrachium rosenbergii, larvae was investigated. Survival rate, weight gain, and larval development were evaluated for different concentrations of nitrate in three experiments. The experiments were divided i n to two phases. In the first phase, larvae from stages I through VIII were analysed, while in the second phase larvae from stage VIII through post-larvae metamorphosis were analysed. Oxygen consumption was also determined for zoea I, II, and VIII exposed to 0, 700, and 1,000 mg/L of nitrate-N. No effect was observed for concentrations up to 180 mg/L NO3-N (experiments I and II), and nitrate levels as 1,000 mg/L NO3-N did not affect survival in the first phase of the third experiment. On the other hand, larval stage index (LSI) and weight gain decreased as nitrate-N concentration increased from 0 to 1,000 mg/L. In the second phase, survival and metamorphosis rate decreased as nitrate concentration increased, according to a linear model. The effect of nitrate levels on weight gain followed a curvilinear pattern. Larval respiration decreased in the water where nitrate was added, but only during stage II. The results demonstrated that nitrate presents extremely low toxicity for giant river prawn larvae, and data were related to the levels of nitrate that usually occur in larviculture systems also discussed. Therefore, nitrate is not a limiting factor for giant river prawn larviculture. © 2003 by The Haworth Press, Inc. All rights reserved.

In vitro antifungal susceptibility of Candida spp. isolates from patients with chronic periodontitis and from control patients

Superinfection by Candida can be refractory to conventional periodontal treatments in specific situations, such as in immunocompromised patients. In these cases, the systemic therapy with antifungal drugs could be indicated. The aim of this study was to analyse antifungal susceptibility of Candida spp. strains isolated from chronic periodontitis patients and from control individuals. A total of 39 C. albicans isolates, 9 C. tropicalis, 2 C. glabrata and 5 Candida spp. from control individuals and 30 C. albicans, 3 C. tropicalis and 2 C. glabrata from periodontitis patients were tested. In the control group, 1 isolate of C. glabrata was resistant to ketoconazole and 1 Candida spp. was resistant to amphotericin B, ketoconazole and miconazole. Among the isolates of periodontitis group, 1 (3.33%) C. albicans isolate was resistant to flucytosine and ketoconazole. According to the obtained results, it could be concluded that fluconazole was the most effective drug against the several Candida species studied. There were not expressive differences in the susceptibility of isolates from periodontitis patients or from control individuals.

Cloning and characterization of the cDNA for the Brazilian Cratomorphus distinctus larval firefly luciferase: Similarities with European Lampyris noctiluca and Asiatic Pyrocoelia luciferases

Studies on firefly (Lampyridae) luciferases have focused on nearctic species of Photinus and Photuris and Euroasiatic species of Lampyris, Luciola, Hotaria, and Pyrocoelia. Despite accounting for the greatest diversity of fireflies in the world, no molecular studies have been carried out on the highly diverse genera from the neotropical region. Here we report the luciferase cDNA cloning for the larva of the Brazilian firefly Cratomorphus distinctus. The cDNA has 1978 bp and codes for a 547-residue-long polypeptide. Noteworthy, sequence comparison as well as functional properties show the highest degree of similarity with Lampyris noctiluca (93%) and Pyrocoelia spp. (91%) luciferases, suggesting a close phylogenetic relationship despite the geographical distance separating these species. The bioluminescence emission spectrum peaks at 550 nm and, as expected, is sensitive to pH, shifting to 605 nm at pH 6. The kinetic properties of the recombinant luciferase were similar to those of other firefly luciferases. © 2004 Elsevier Inc. All rights reserved.

Larval development of the giant river prawn Macrobrachium rosenbergii at different ammonia concentrations and pH values

The effect of ammonia and pH levels on giant river prawn Macrobrachium rosenbergii larvae were evaluated to provide science-based information on safe levels of ammonia and pH for larviculture. Survival rate, developmental stage, and larval weight gain were determined for larvae kept in water with total ammonia (NH4-N) concentrations of 0, 1, 2, 4, and 8 mg\L and pH 7, 8, and 9. The trials were conducted in two phases: phase 1, larvae from stages I through VIII and phase 2, larvae from stage VIII until metamorphose. Oxygen consumption was determined for larvae in stages I and VIII at total ammonia concentrations of 0, 4, and 8 mg/L and pH 8. Survival rate up to stage VIII varied from 86 to 98% and did not differ for total ammonia concentrations in pH 7 and 8 and for 0 mg/L NH4-N in pH 9. Survival rate was significantly lower (0-20%) for total ammonia concentrations from 1 to 8 mg/L (0.43-3.41 mg/L of unionized ammonia) in pH 9. Larval stage indexes (7.9-8.0 range) and weight gain (1.572-2.931 mg range) of larvae at the end of phase 1 of the experiment did not differ for the different ammonia concentration solutions, but were significantly lower in pH 9. In phase 2, no parameter differed among treatments for pH 7 and 8; however there was total mortality at pH 9 until 96h. Respiration rates diminished when larvae were exposed to total ammonia concentrations of 4 and 8 mg/L (0.28 and 0.55 mg/L of unionized ammonia), but development remained unaltered. Therefore, M. rosenbergii larvae tolerate high levels of total ammonia, while toxicity depends primarily on unionized ammonia concentrations. In addition, alkaline pH (9) acted directly on the larvae, curbing development and causing severe mortality. Larval tolerance to high ammonia and pH levels decreases for the last zoeal stages. © Copyright by the World Aquaculture Society 2005.

Analysis of susceptibility profile of Pseudomonas spp. and prevalence of bacterial samples from the surfaces of dental consulting-rooms

The aim of this research was to evaluate the susceptibility profile of Pseudomonas spp. and the prevalence of bacterial samples isolated from horizontal surfaces surrounding wash-basins used by dentists in several adjoined consulting-rooms, at points next to and at a distance from the basin, before and after surgical procedures. Our results showed a high percentage of Gram-positive cocci and Gram-negative bacilli; 34.66% were Staphylococcus spp. and 30.12% were non-fermentative Gram-negative bacilli among which Pseudomonas spp. (40.90%) was the commonest genus. Analysis of the susceptibility profile of Pseudomonas spp. isolates by determining the minimal inhibitory concentration (MIC) of 14 antibiotics showed a great variation among the strains and high rates of resistance to cefazolin, ceftazidime and aztreonan. Of the 14 antibiotics tested, 59.03% were found to be active against all the environmental isolates. Strains were resistant to aztreonan (62.82%), while susceptibility to third generation cephalosporins was variable.

Evaluation of the antimicrobial susceptibility of methicillin-resistant Staphylococcus aureus isolated from human infections

Multiresistant Staphylococcus aureus constitutes an important public health problem, especially in view of its possible spread in nosocomial environments. In the present work, we analyzed the susceptibility profile of 80 S. aureus stains from human infections resistant to at least 10 drugs. For this study, the techniques used were the disk method and minimum inhibitory concentration (MIC) of the following drugs: cefuroxime, ciprofloxacin, clindamycin, erythromycin, gentamycin, imipenem, oxacillin, rifampicin, tetracycline and vancomycin, according the criteria of the National Committee for Clinical Laboratory Standards (NCCLS). Methicillin was included in the antibiogram as a marker, which is usually used in drugs selection for the treatment of staphylococcal infections. Results indicated that the most effective drug was vancomycin. For the other 10 drugs, the percentage of resistant strains ranged from 85% to 93.75%. In relation to the MICs, it was observed that vancomycin (MIC 90% = 0.615ug/ml) was the most effective drug; followed by rifampicin (MIC 90% = 2.6ug/ml) and ciprofloxacin (MIC 90% = 26.6ug/ml). The drugs that showed the least effective activity were cefuroxime, clindamycin, erythromycin, gentamycin, and oxacillin. On the other hand, observation of β-lactamase production revealed that most of the methicillin-resistant strains produced β-lactamase (83.7%), potentiating the risks of nosocomial infections. In general, vancomycin still continues to be one of the most effective drugs for staphylococcal infections therapy.