Fecal Droplets Research Articles

Relato de coccidiose em um trinca-ferro Saltator similis de vida-livre no Parque Nacional do Itatiaia no sudeste brasileiro

ABSTRACT: The green-winged saltator Saltator similis (d’Orbigny & Lafresnaye, 1837) is a passerine bird highly valued by bird breeders and a target of illegal wildlife trafficking. Coccidian chromists are etiological agents of coccidiosis, which may be asymptomatic and enzootic in bird populations when environmental conditions are favorable. However, epizootic outbreaks with severe disease can occur due to environmental changes. In this context, this study reports a case of coccidiosis in a free-living juvenile specimen of S. similis captured at the Itatiaia National Park in southeastern Brazil. Typical clinical signs of coccidiosis were observed, and it was associated with greenish mucoid diarrhea containing a high density of coccidian oocysts. Three Isospora spp. were specifically identified in 14 fecal samples (fecal droplets) collected over two hours. Isospora saltatori (Berto, Balthazar, Flausino & Lopes, 2008) had the highest density and positivity throughout most of the collection period. Finally, this paper discusses the importance of the continuous evaluation of the health of birds as bioindicators and their coccidian ecological biomarkers as one of the strategies for evaluating the conservation status of Itatiaia National Park.

Active Surveillance and Genetic Characterization of Prevalent Velogenic Newcastle Disease and Highly Pathogenic Avian Influenza H5N8 Viruses Among Migratory Wild Birds in Southern Egypt During 2015-2018.

A total of 1007 samples (910 fecal droplets and 97 cloacal swabs) were collected from 14 species of migratory wild birds in most wetlands during 3 successive migration seasons from September to March (2015-2018) in Southern Egypt. The samples were propagated inembryonated chicken eggs and positive allantoic fluids by hemagglutination test were tested for Newcastle disease virus (NDV) and avian influenza virus (AIV) prevalence using RT-PCR and specific primers targeting the NDV fusion (F) and AIV matrix genes. Further subtyping of the AIV hemagglutinin (HA) and neuraminidase (NA) was conducted, and representative isolates were selected and sequenced for full F gene of NDVs and HA and NA genes of the AIV. Overall isolation rate of hemagglutinating viruses was 5.56% (56/1007), from them 5.36% (3/56) AIV, 85.71% (48/56) NDV and 8.93% (5/56) co-infection of NDV and AIV was detected. The sequences analysis of full F genes of 10 NDV isolates revealed that they have multi-basic amino acid motifs 111E/GRRQKR/F117 as velogenic strains with nucleotides and amino acids similarities of 96-100%. In addition, they phylogenetically clustered into groups and subgroups within genotype VII.1.1 and sub-genotype VIIj with a close relation to NDVs isolated from chickens in Egypt. The AIVH5N8 subtype was in clade 2.3.4.4b with a highly pathogenic nature and close relation to Egyptian domesticated H5N8 viruses rather than those from wild birds. The current data showed the contribution of migratory birds to the continuous circulation of virulent NDV and AIVH5N8 among domesticated chickens in Southern Egypt.

Open defecation and squat toilets, an overlooked risk of fecal transmission of COVID-19 and other pathogens in developing communities.

The novel coronavirus disease COVID-19 has infected over 46 million people in 219 countries and territories. Following evidence of viral loadings and infectivity of feces of infected individuals, public health authorities have suggested to take precautions on the transmission of COVID-19 via fecal-associated routes. Recent discussions on fecal transmission of COVID-19 have mainly focused on municipal sewage. Yet, a widely neglected aspect in containing the virus is that a major part of the population in developing regions do not have access to private, clean sanitary facilities. Therefore, we hypothesize that open defecation and the prevalent use of squat toilets are additional risk factors in those communities. Here, we review fecal transmission of COVID-19, the practices of open defecation, and the resultant routes of transmission of fecal pathogens. Also, we highlight the open design of common squat toilets and the potential exposure to fecal droplets and residues. We observed that at least 20 countries reporting more than 10,000 confirmed infections have 5–26% of their population practicing open defecation. We illustrate the potential routes of transmission of COVID-19 and other fecal pathogens via human feces in communities practicing open defecation. Here, poor hand hygiene, contaminated shoes and objects, mechanical vectors, and outdoor human activities can all contribute to fecal transmission. Other risk factors include squat pans with lidless designs and open flushing mechanisms, in-cubicle open waste bins, and the lack of water-sealing U-traps in squat toilets.

SARS-CoV-2: The Growing Case for Potential Transmission in a Building via Wastewater Plumbing Systems.

SARS-CoV-2: The Growing Case for Potential Transmission in a Building via Wastewater Plumbing Systems.

Enrichment and Broad Representation of Plant Biomass-Degrading Enzymes in the Specialized Hyphal Swellings of Leucoagaricus gongylophorus, the Fungal Symbiont of Leaf-Cutter Ants.

Leaf-cutter ants are prolific and conspicuous constituents of Neotropical ecosystems that derive energy from specialized fungus gardens they cultivate using prodigious amounts of foliar biomass. The basidiomycetous cultivar of the ants, Leucoagaricus gongylophorus, produces specialized hyphal swellings called gongylidia that serve as the primary food source of ant colonies. Gongylidia also contain plant biomass-degrading enzymes that become concentrated in ant digestive tracts and are deposited within fecal droplets onto fresh foliar material as ants incorporate it into the fungus garden. Although the enzymes concentrated by L. gongylophorus within gongylidia are thought to be critical to the initial degradation of plant biomass, only a few enzymes present in these hyphal swellings have been identified. Here we use proteomic methods to identify proteins present in the gongylidia of three Atta cephalotes colonies. Our results demonstrate that a diverse but consistent set of enzymes is present in gongylidia, including numerous plant biomass-degrading enzymes likely involved in the degradation of polysaccharides, plant toxins, and proteins. Overall, gongylidia contained over three quarters of all biomass-degrading enzymes identified in the L. gongylophorus genome, demonstrating that the majority of the enzymes produced by this fungus for biomass breakdown are ingested by the ants. We also identify a set of 40 of these enzymes enriched in gongylidia compared to whole fungus garden samples, suggesting that certain enzymes may be particularly important in the initial degradation of foliar material. Our work sheds light on the complex interplay between leaf-cutter ants and their fungal symbiont that allows for the host insects to occupy an herbivorous niche by indirectly deriving energy from plant biomass.

Bed Bug Detection: Current Technologies and Future Directions

Technologies to detect bed bugs have not kept pace with their global resurgence. Early detection is critical to prevent infestations from spreading. Detection based exclusively on bites is inadequate, because reactions to insect bites are non-specific and often misdiagnosed. Visual inspections are commonly used and depend on identifying live bugs, exuviae, or fecal droplets. Visual inspections are inexpensive, but they are time-consuming and unreliable when only a few bugs are present. Use of a dog to detect bed bugs is gaining in popularity, but it can be expensive, may unintentionally advertise a bed bug problem, and is not foolproof. Passive monitors mimic natural harborages; they are discreet and typically use an adhesive to trap bugs. Active monitors generate carbon dioxide, heat, a pheromone, or a combination to attract bed bugs to a trap. New technologies using DNA analysis, mass spectrometry, and electronic noses are innovative but impractical and expensive for widespread use.

Leaf-cutting ant fungi produce cell wall degrading pectinase complexes reminiscent of phytopathogenic fungi

BackgroundLeaf-cutting (attine) ants use their own fecal material to manure fungus gardens, which consist of leaf material overgrown by hyphal threads of the basidiomycete fungus Leucocoprinus gongylophorus that lives in symbiosis with the ants. Previous studies have suggested that the fecal droplets contain proteins that are produced by the fungal symbiont to pass unharmed through the digestive system of the ants, so they can enhance new fungus garden growth.ResultsWe tested this hypothesis by using proteomics methods to determine the gene sequences of fecal proteins in Acromyrmex echinatior leaf-cutting ants. Seven (21%) of the 33 identified proteins were pectinolytic enzymes that originated from the fungal symbiont and which were still active in the fecal droplets produced by the ants. We show that these enzymes are found in the fecal material only when the ants had access to fungus garden food, and we used quantitative polymerase chain reaction analysis to show that the expression of six of these enzyme genes was substantially upregulated in the fungal gongylidia. These unique structures serve as food for the ants and are produced only by the evolutionarily advanced garden symbionts of higher attine ants, but not by the fungi reared by the basal lineages of this ant clade.ConclusionsPectinolytic enzymes produced in the gongylidia of the fungal symbiont are ingested but not digested by Acromyrmex leaf-cutting ants so that they end up in the fecal fluid and become mixed with new garden substrate. Substantial quantities of pectinolytic enzymes are typically found in pathogenic fungi that attack live plant tissue, where they are known to breach the cell walls to allow the fungal mycelium access to the cell contents. As the leaf-cutting ant symbionts are derived from fungal clades that decompose dead plant material, our results suggest that their pectinolytic enzymes represent secondarily evolved adaptations that are convergent to those normally found in phytopathogens.

EPHEMERAL WINDOWS OF OPPORTUNITY FOR HORIZONTAL TRANSMISSION OF FUNGAL SYMBIONTS IN LEAF-CUTTING ANTS

Evolutionary theory predicts that hosts are selected to prevent mixing of genetically different symbionts when competition among lineages reduces the productivity of a mutualism. The symbionts themselves may also defend their interests: recent studies of Acromyrmex leaf-cutting ants showed that somatic incompatibility enforces single-clone gardens within mature colonies, thereby constraining horizontal transmission of fungal symbionts. However, phylogenetic analyses indicate that symbiont switches occur frequently enough to remove most signs of host-symbiont cocladogenesis. Here we resolve this paradox by showing that transmission among newly founded Acromyrmex colonies is not constrained. All tested queens of sympatric A. octospinosus and A. echinatior offered a novel fragment of fungus garden accepted the new symbiont. The outcome was unaffected by genetic distance between the novel and the original symbiont, and by the ant species the novel symbiont came from. The colony founding stage may thus provide an efficient but transient window for horizontal transmission, in which the fungus is unable to actively defend its partnership position before the host feeds on it, so that host fecal droplets remain compatible with alternative strains during the early stage of colony founding. We discuss how brief stages of low commitment between partners may increase the evolutionary stability of ancient coevolved mutualisms.

Excretion of Phosphoric Acid by Red Imported Fire Ants,Solenopsis invictaBuren (Hymenoptera: Formicidae)

Food sources, mineral redistribution, and excreta are some of the ways ants enrich their mound environment. Phosphorus excretion has not been reported in red imported fire ants, Solenopsis invicta Buren. Quantification of phosphoric acid in worker fecal droplets and larval anal liquid of S. invicta was performed. Phosphoric acid was trimethylsilylated using N,O-bis(trimethylsilyl)trifluoroacetamide, and the tris(trimethylsilyl) phosphate was analyzed using gas chromatography-mass spectrometry. Phosphoric acid made up 2.13 ± 1.08% (SD) of the dry weight of worker fecal droplets and 2.09 ± 0.72% of the larval anal liquid. The rate of phosphoric acid production in workers was 0.08 ± 0.024 μg/ant/d. Because the hindgut of red imported fire ant larva is separated from the midgut, the excretory products in larval anal liquid are primarily of metabolic origin. The presence of phosphoric acid in larval anal liquid shows that red imported fire ants excrete phosphoric acid.

Excretion of Phosphoric Acid by Red Imported Fire Ants, <I>Solenopsis invicta</I> Buren (Hymenoptera: Formicidae)

Food sources, mineral redistribution, and excreta are some of the ways ants enrich their mound environment. Phosphorus excretion has not been reported in red imported fire ants, Solenopsis invicta Buren. Quantification of phosphoric acid in worker fecal droplets and larval anal liquid of S. invicta was performed. Phosphoric acid was trimethylsilylated using N,O-bis(trimethylsilyl)trifluoroacetamide, and the tris(trimethylsilyl) phosphate was analyzed using gas chromatography-mass spectrometry. Phosphoric acid made up 2.13 ± 1.08% (SD) of the dry weight of worker fecal droplets and 2.09 ± 0.72% of the larval anal liquid. The rate of phosphoric acid production in workers was 0.08 ± 0.024 μg/ant/d. Because the hindgut of red imported fire ant larva is separated from the midgut, the excretory products in larval anal liquid are primarily of metabolic origin. The presence of phosphoric acid in larval anal liquid shows that red imported fire ants excrete phosphoric acid.

Triatoma dominicana sp. n. (Hemiptera: Reduviidae: Triatominae), and Trypanosoma antiquus sp. n. (Stercoraria: Trypanosomatidae), the First Fossil Evidence of a Triatomine-Trypanosomatid Vector Association

Triatoma dominicana sp.n. (Hemiptera: Reduviidae: Triatominae) is described from a fifth instar nymph exuvia in Dominican amber and represents the first fossil representative of this subfamily. The new species is placed in the tribe Triatomini and tentatively in the genus Triatoma on the basis of the following characters; body length of 19.5 mm, body and appendages without long hairs, antenniferous tubercles without conspicuous apicolateral projections; abdomen with a series of five median tubercles; only fourth antennal segment annulated; lateral postocular callosities absent; stridulatory sulcus present and pronotum with discal and lateral tubercles. Trypanosoma antiquus sp. n. is described from two fecal droplets adjacent to Triatoma dominicana, thus providing the first fossil evidence of a triatomine-trypanosomatid vector association. The metatrypanosome stages of the fossil are similar in size to those of T. cruzi and could be a progenitor of this species. Mammalian hairs adjacent to the fecal droplets suggest that the vertebrate host for both the triatomine and trypanosomatid was a bat. Based on these findings, it is possible that bats were the original vertebrate hosts of T. cruzi-like trypanosomes.

Mutualistic Fungi Control Crop Diversity in Fungus-Growing Ants

Leaf-cutting ants rear clonal fungi for food and transmit the fungi from mother to daughter colonies so that symbiont mixing and conflict, which result from competition between genetically different clones, are avoided. Here we show that despite millions of years of predominantly vertical transmission, the domesticated fungi actively reject mycelial fragments from neighboring colonies, and that the strength of these reactions are in proportion to the overall genetic difference between these symbionts. Fungal incompatibility compounds remain intact during ant digestion, so that fecal droplets, which are used for manuring newly grown fungus, elicit similar hostile reactions when applied to symbionts from other colonies. Symbiont control over new mycelial growth by manurial imprinting prevents the rearing of multiple crops in fungus gardens belonging to the same colony.

Nest-founding in Acromyrmex octospinosus (Hymenoptera, Formicidae, Attini): demography and putative prophylactic behaviors

Foundresses of the leaf-cutting ant Acromyrmex octospinosus in central Panama forage for leaves as garden substrate (semi-claustral foundation). The fungal pellet and substrate usually are attached to rootlets, which are used as a platform for the garden. This arrangement keeps the garden suspended away from the earthen chamber of the underground nest during early colony growth, and we hypothesize that it serves to minimize contact between the garden and contaminants. A. octospinosus foundresses produce from 3 to 7 workers in 2.7 months after founding the nest, but workers do not forage for substrate at this time. Incipient nests died or were abandoned at a monthly rate of ca. 50%. We show that ants routinely clean their legs before manipulating the garden substrate. We also describe how foundresses use their fore-legs to rub the surface of the metapleural gland (MPG), and they then use typical grooming behaviors to pass the forelegs through the mouthparts, after which the ant then licks the garden substrate. Similarly, ants apparently use their mouths to transfer fecal droplets to their legs. We briefly discuss the functional significance of these grooming behaviors, and hypothesize that they are prophylactic behaviors that may help the foundress maintain a hygienic garden.

Predation behaviors of Franklinothrips orizabensis (Thysanoptera: Aeolothripidae) towards Scirtothrips perseae and Heliothrips haemorrhoidalis (Thysanoptera: Thripidae)

The foraging behaviors of predatory female Franklinothrips orizabensis Johansen towards Scirtothrips perseae Nakahara and Heliothrips haemorrhoidalis (Bouch e in avocado leaf arenas were videotaped and analyzed. F. orizabensis encountered and attacked more second instar S. perseae with 80% ðn ¼ 113Þ attacks on this life stage being observed compared to 20% on first instars ðn ¼ 28Þ when equal numbers of these prey were presented simultaneously. However, there was no significance difference by life stage for the probability of being attacked after being encountered. Successful attacks by F. orizabensis only occurred against propupal H. haemorrhoidalis ðn ¼ 6Þ that lacked protective fecal droplets, 65% of prey encounters were with this life stage, and 6% of encounters with propupae resulted in attacks. Second instar H. haemorrhoidalis larvae bearing protective droplets were encountered less frequently and were not attacked when contacted even though equal numbers of both stages were presented simultaneously. Consequently, the mean proportion of the time spent grooming by F. orizanbensis females attacking immature H. haemorrhoidalis was four times greater than that for females attacking S. perseae larvae even though there was no significant difference in mean time (s) spent in each grooming bout. Significant differences in mean consumption times by F. orizabensis existed across life stages and prey species. Consumption times were shortest for first instar S. perseae larvae, intermediate for second instars, and longest for H. haemorrhoidalis propupae. Mean consumption times for sequentially attacked second instar S. perseae declined significantly indicating satiation of predators. Predators would spend on average approximately 7–13 s probing leaves with mouthparts and feeding on fluids; a behavior that on average occupied 2–5% of their time. This result may explain observed mortality of F. orizabensis in the laboratory and field when this predator forages on avocado leaves that have been treated with insecticides that exhibit translaminar activity. 2003 Elsevier Science (USA). All rights reserved.

Stercorarial shedding and transtadial transmission of hepatitis B virus by common bed bugs (Hemiptera: Cimicidae).

Transtadial persistence and stercorarial shedding of hepatitis B virus (HBV) in common bed bugs, Cimex lectularius L., was studied by using experimental infectious blood feedings, infectious intrathoracic inoculations, and virus detection by polymerase chain reaction and Southern hybridization. Results showed that HBV persisted after an infectious blood meal in bed bug bodies for up to 35 d after the infectious blood meal. It was passed transtadially through one molt regardless of instar, was shed in fecal droplets for up to 35 d after the infectious blood meal, but was not passed transovarially. In bugs inoculated intrathoracically, HBV was detected for 21 d postinoculation. Previous studies detected the hepatitis B surface antigen found on both infectious and noninfectious particles in bed bugs. In this study, the presence of nucleic acids amplified from a conserved core region of the viral genome in bodies and feces of C. lectularius suggests that the HBV virus may be mechanically transmitted in feces or when bugs are crushed, during feeding.

Transmission Mechanisms ofNosema empoascae(Microspora: Nosematidae) inEmpoasca fabae(Homoptera: Cicadellidae)

Four experiments investigating horizontal and vertical transmission mechanisms of the microsporidiumNosema empoascae,in a population of the potato leafhopper,Empoasca fabae,were conducted. Transovarial transmission was the only route of transmission demonstrated experimentally. Transmission on the egg surface was unlikely. Spores were not present in fecal droplets, and in laboratory experiments the microsporidium could not be transmitted horizontally, either by contact or in venereal transmission experiments. Thus, this microsporidium persists primarily, if not exclusively, by transovarial transmission in the leafhopper populations. If horizontal transmission occurs, it occurs at a very low rate through an as yet undetermined mechanism. The ramifications of transovarial transmission as an exclusive mechanism are discussed.

Biology and ecology of bat flies (Diptera: Streblidae) on bats in the genus Carollia.

The biology and ecology of bat flies found on Carollia perspicillata were investigated in Heredia Province, Costa Rica. Only Trichobius joblingi, Speiseria ambigua , and Strebla guajiro were found on Carollia . These streblids were absent from 9 species of bats in 7 other genera. Although streblids transfer readily among bats in a roost, flies had a clumped distribution on foraging bats. Female bats had significantly higher numbers of flies (3.5 ± 3) than did male bats (2.1 ± 2). The ratio of female to male flies was 1:1.3, 1:1.4, and 1:1.7 for T. joblingi, Sp. ambigua , and St. guajiro , respectively. Pupiposition occurred between 1730–1930 h, coinciding with the time when bats leave their roosts. Pupae eclosed during the morning, providing teneral flies with maximum time for cuticle hardening and host searching. Mean pupal developmental times (in days) for caged T. joblingi, Sp. ambigua , and St. guajiro were 17.4 ± 1, 19.8 ± 0.66, and 18.8 ± 0.75, respectively; pupal mortality was 22, 48, and 62%, respectively. Trichobius joblingi pupae monitored in a tree roost had 37% mortality. Streblids fed frequently and voided fecal droplets containing masses of intact red blood cells. When removed from hosts, fly defecation rates dropped to 0 after 1 h. Fed T. joblingi had a mean survivorship off the host of 2.79 ± 0.88 h, whereas teneral flies lived significantly longer (4.0 ± 0.47 h). Mean survival time off the host for Sp. ambigua and St. guajiro was less than 7 h. Streblids copulated on the host, and males would often “ride” fully parous females. Laboulbeniomycete fungi were found on all 3 bat fly species; this is the first record of them on New World streblids.

Effect of ultraviolet radiation on the microsporidian Octosporea muscaedomesticae with reference to protectants provided by the host Phormia regina

The effect of ultraviolet light on the microsporidian Octosporea muscaedomesticae in relation to infection in the adult black blowfly, Phormia regina, was investigated. A 30-Watt germicidal lamp, 253.7-nm wavelength, was used as source of uv light in five investigations. In addition, sunlight served as a uv source in two studies. Viable naked dried spores exposed to the uv lamp at a distance of 10 cm were killed after 15 min. Viable naked spores in an aqueous suspension were killed after 30 min of exposure to the uv lamp and after 3 hr of exposure to bright sunlight, respectively. Daily 30-min uv lamp exposures on living hosts harboring all life phases of the parasite did not interfere with the ensuing infection in the blowfly's midgut and the pathogen's developmental cycle. Spores harvested from uv-treated infected hosts were found to be as infective as spores retrieved from hosts not treated with uv. Spores contained in dried fecal droplets and exposed up to 3 hr to the uv lamp, or 12 hr to bright sunlight, respectively, remained infective. Addition of uric acid to a preparation of naked spores prior to 15- and 30-min uv irradiations yielded 100% infection in both host groups. A uv-protective function is ascribed to components provided by the host's tissues and feces.

A potential infectivity index for Triatoma harboring Trypanosoma cruzi Chagas

The contaminative effect of conenose bugs, Triatoma protracta, for man and animals is greater for nymphs than adults in relation to number of defecations released over a 6-hour observation period. An infectivity index is proposed for the number of metacyclic parasites present in proportion to non-metacyclic per fecal droplet. The total contaminative potential averaged 5.6 fecal droplets released per nymph and 2.6 per adult. The total number of parasites per bug averaged 555.8 for 19 different samples from 15 adults and 157.5 for samples from 50 nymphs. Time of first defecation favors 5th instar nymphs for contaminative effect nearest to host immediately after feeding. The number of metacyclic Trypanosoma cruzi in 1st fecal droplets released averaged 349.2 per female, 156.8 per male and 154.1 per 4th instar nymph. The total number of metacyclic parasites per bug with complete counts averaged 1954.3 per male (9 ♂o ♂o), 1821.1 per female (6 ♀o ♀o) and 1678 per 4th instar nymph (4 bugs) 34 to 1043 days after the infective blood meal.

Febre maculosa das Montanhas Rochosas: ensaios negativos de transmissão experimental do virus por Triatomideos

1. - As seguintes especies de Triatomideos não puderam transmitir pela picada o virus da febre maculosa das Montanhas Rochosas a cobayas normaes, nos seguintes prazos após a sucção de animal infectado: Eutriatoma uhleri, 33, 47, 75 e 141 dias (1 exemplar); Triatoma protracta, 15 e 37 dias (1 exemplar), Triatoma infestans, 8 dias (15 exemplares), e Rhodnius prolixus, 2 dias (1 exemplar). Foi demonstrado por inoculaçao que o ultimo insecto ainda continha o virus vivo. 2. - Experiencias de transmissão mechanica, por picada interrompida em animal infectado e continuada immediatamente em animal são, foram tambem negativas, com as especies T. protracta e R. prolixus. Um unico insecto da primeira especie picou 2 vezes cada animal, emquanto que 22 exemplares da segunda especie picaram de 1 a 3 vezes cada cobaya. 3. - A inoculação de gottas de dejecções de um R. prolixus eliminadas 2 dias depois de sugar animal infectado, não produziu a infecção em cobaya normal, não obstante ter sido demonstrada a presença do virus no organismo do barbeiro, por inoculação do conteúdo estomacal em outra cobaya. 4. - Foram feitas experiencias para determinar o tempo de sobrevivencia do virus nos barbeiros, inoculando-se conteúdo intestinal a diversos intervallos depois da sucção de cobayas infectadas, com os seguintes resultados: T. infestans: positivo 1 vez em 24 horas e 2 vezes em 48 horas; negativo 2 vezes em 72. 96, 120 e 192 horas. P. megistus: positivo 3 vezes em 24 horas, 2 vezes em 48 horas e 1 vez em 72 horas; negativo 1 vez em 72 e 96 horas; resultado duvidoso ou sem valor (infecção intercorrente) 1 vez em 48 horas e 2 vezes a 72, 96 e 144 horas cada um. R. prolixus: positivo 1 vez em 24, 48 e 72 horas e negativo em 96 horas. 5. - Em vista dos resultados destas experiencias, feitas com 5 especies pertecentes a 4 generos de Triatomideos, torna-se muito pouco provavel que estes Hemipteros possam transmitir pela picada o virus da febre maculosa das Montanhas Rochosas, ou retel-o em seu organismo, em estado virulento, por mais de 2 a 4 dias.