Uso de alguns estimadores ridge na análise estatística de experimentos em entomologia

Inúmeros experimentos em ciências agrárias apresentam variáveis que podem dar origem a problemas de multicolinearidade. Em se tratando da aplicabilidade de modelos de regressão, o problema da multicolinearidade tem como principal consequência o inflacionamento dos erros padrão e, com isso, o valor da estatística t-student é reduzido de tal forma que interfere nos resultados inferenciais. Várias medidas são propostas, na literatura, para resolver o problema de multicolinearidade. Entretanto, o desempenho dessas medidas está sujeito ao grau de multicolinearidade que as variáveis poderão apresentar, bem como ao tamanho amostral. Frente a este problema, este trabalho tem por objetivo avaliar alguns estimadores ridge, utilizando simulação Monte Carlo, bem como, apresentar a aplicação desses estimadores em um experimento, com dados reais, na área de entomologia. Mediante esta aplicação, os resultados expressivos alcançados foram obtidos em função da eficiência dos estimadores ridge avaliados, em relação ao estimador de mínimos quadrados. Em se tratando dos resultados computacionais, concluiu-se que estimadores ridge avaliados são recomendáveis, em experimentos que considerem as variáveis com diferentes graus de multicolinearidade, para amostras maiores do que n=50.

Modelos ridge em planejamentos de misturas: uma aplicação na extração da polpa de pequi

Mixture Models can be used in experimental situations involving areas related to food science and chemistry. Some problems of a statistical nature can be found, such as effects of multicollinearity that result in uncertainty in the optimization of a dependent variable. This study proposes the application of the ridge model adapted for mixture planning considering the Kronecker (K-model) and Scheffe (S-Model) methods applied to response surfaces. The method determined the proportions of hexane, acetone and alcohol proportions that resulted in the maximum response of percentage of extracted pequi (Caryocar brasiliense) pulp oil.

Characterization of heat tolerance in wheat cultivars and effects on production components

Abstract: There is a need for heat tolerant wheat cultivars adapted to the expansion of cultivation areas in warmer regions due to the high demand of this cereal for human consumption. The objective of this study was to evaluate the effect of high temperatures on grain yield and yield components of wheat and characterize heat tolerant wheat genotypes at different development stages. The genotypes were evaluated in the field with and without heat stress. High temperatures reduced the number of spikelets per spike (21%), number of grains per spike (39%), number of grains per spikelet (23%), 1000-grain weight (27%) and grain yield (79%). Cultivars MGS 1 Aliança, Embrapa 42, IAC 24-Tucuruí and IAC 364-Tucuruí III are the most tolerant to heat stress between the stages double ridge and terminal spikelet; MGS 1 Aliança, BRS 264, IAC 24-Tucuruí, IAC 364-Tucuruí III and VI 98053, between meiosis and anthesis; and BRS 254, IAC-24-Tucuruí, IAC-364-Tucuruí III and VI 98053, between anthesis and physiological maturity. High temperatures reduce grain yield and yield components. The number of grains per spike is the most reduced component under heat stress. The genotypes differed in tolerance to heat stress in different developmental stages.

Anfíbios Anuros da coleção Adolpho Lutz: III - Hyla claresignata Lutz & B. Lutz, 1939

Hyla claresignata Lutz & Lutz, 1939, is a large species apparently not closely allied to the other known Brazilian hylas. It is characterized by the very small tympanum; the head is short and the snout rounded; the legs are long, the hands and feet unusually large, the latter extensively webbbed. The specific name is derived from the insular, irregular, or roughly triangular, dark spots, with a light halo, found mostly in the dorso-lateral region and on the legs. It belongs to the rain-forest fauna of the Marítime Range. The adult is a bromeliad-dweller and the tadpole rhyacophilous. DESCRIPTION. Vomerine teeth in two separate, oblique, groups, behind the large choanae, parallel to the posterior half of their inner border. Tongue entire, short, very broad and hardly free behind. Snout short, rounded, with distinct canthus rostralis and gradually sloping loreal region. Eye very large and prominent, its horizontal diameter almost equal to the distance between its anterior corner and the tip of the snout. Tympanum very small, less than one third of the diameter of the eye, but distinct, partly covered by a short, heavy ridge. Lateral fingers less than one third webbed; fourth finger slightly longer than the second, just reaching the base of the disk of the third; subarticular tubercles well developed; an angular pollex rudiment, more noticeable in the males. Toes almost completely webbed, the edge of the web inserted at the base of the disk on the third and the fifth; an inner metatarsal tubercle. Skin smooth above, granular beneath, on the throat minutely so. No dermal appendage on the hell. Habit robust, head broader than long, body rather heavy, slightly narrowed in the postaxillary region. Legs long, the tibiotarsal articulation reaching beyond the tip of the snout when adpressed. Type (female): 61 mm. (Fig. 1.) DIAGNOSIS of TADPOLE (by G. Orton). "A large specialized, mountain-stream tadpole, with wide head an elongated, flattened snout, greatly enlarged lips and high tooth formula. Eyes dorsal. Spiracle sinistral, projecting, situated far back on side. Anus dextral. Tooth formula 8/12 to 9/14 in fully grown larvae. Tail with a prominent, vertical dark band across musculature and fins; a second concentration of dark pigment near tip of tail, may or may not form a similar but narrower band. Maximum known total length: 60mm.; head and body length 25mm. (Figs. 6 e 7). For further details see Lutz & Lutz, 1939 and Lutz B. & Orton G. 1946.

Observation on the morphology of Australorbis glabratus

The present morphological study of A. glabratus was based on the observation of shell, radula, renal region and genitalia of 50 specimens having a shell diameter of 18 mm. In this summary we record the data pertaining to the chracteristics that can be used in systematics. The numerals refere to the mean and their standard deviation; no special reference being made, they correspond to length measurements. Shell: 18 mm in diameter, 5.59 ± 0.24 mm in greatest width, 5 to 6 whorls. Right side umbilicated, left one weakly depressed. Last whorl about thrice as tall as the penultimate one at the aperture, the measurements being taken on the right side. Aperture perpendicular or a little oblique. Body, extended: 47.06 ± 3.31 mm. Renal tube: Narrow and elongated, 23.84 ± 1.90 mm, showing a pigmented ridge along its ventral surface. Ovotestis: 12.78 ± 1.50 mm. Mainly trifurcate diverticula attaching in fan-like manner to the collecting canal (this arrangement is seen to best advantage in the cephalic middle of the ovotestis). The collecting canal greatly swells at the cephalic end, narrowing suddenly as it leaves the ovotestis. Ovisperm duct: 13.70 ± 1.68 mm, including the non-unwound seminal vesicle. The latter, situated about 1 mm from the beginning af the ovisperm duct, was 1.14 ± 0.29 mm in greatest diameter, and is beset by numerous short diverticula. Sperm duct: 14.16 ± 1.27 mm, pursuing a sinous course along the oviduct. Prostate: Prostate duct 5.53 ± 0.74 mm, collecting a row of long diverticula, the latter 21.6 ± 3.5 in number. Last diverticulum generally simple or bifurcate, penultimate generally arborescent, bifurcate or simple, antepenultimate nearly always arborescent, the remaining ones arborescent. The arborescent diverticula frequently give off secondary branches. Vas deferens: 17.50 ± 2.05 mm. The ratio vas deferens/vergic sac was 4.7 ± 0.6. Verge: 3.70 ± 0.54 mm long, 0.12 ± 0.03 mm wide. Free end tapering to a point where the sperm canal opens. No penial stylet. Vergic sac: 3.77 ± 0.50 mm long, 0.19 ± 0.01 mm wide. The length ratio vergic sac/preputium was 1 ± 0.02. Preputium: Deeply pigmented, 3.79 ± 0.40 mm long, 0.89 ± 0.12 mm wide in the middle. Muscular diaphragm between it and the vergic sac. Two muscular pilasters along its lateral walls. Oviduct: 10.24 ± 1.29 mm, suddenly swollen at the cephalic end so that it forms a folded pouch capping the beginning of the uterus. Uterus: 10.58 ± 1.18 mm. Vagina: 2.06 ± 0.15 mm long, 0.32 ± 0.05 mm wide, showing a swelling at its caudal portion, just above the opening of the spermathecal duct. Spermatheca: 1.57 ± 0.41 mm long, 0.92 ± 0.23 mm wide. Spermathecal duct 1.15 ± 0.23 mm. Radula: 125 to 163 rows of teeth (mean 141.4 ± 9.8). Radula formula 27-1-27 to 34-1-34 (mean 30.9 ± 1.7).

Observations on the morphology of Australorbis nigricans

A morphological study was done on A. nigricans, based on the observation of shell, radula, renal region and genitalia of 50 specimens measuring 18 mm in diameter. The data obtained are to be compared with those recorded in our previous paper (PARAENSE & DESLANDES, 1955) on A. glabratus. The characteristics common to both species will not be mentioned here. The numerals refere to the means and their standard deviations: no special reference being done, they correspond to length measurementes. Shell - 18 mm in diameter, 6.37 ± 0.29 mm in greatest width, 6 whorls. Prevailing colur ferruginous sepia, a minority of olivaceous, ochreous, nigrescent and deeply black specimens being found. Right side variously depressed, umbilicated, 1.5 to 3.5 mm deep from the bottom of the umblicus to the highest level of the last whorl. Left side more depressed than the right one, broadly concave, 1.5 to 3.5 mm deep. Both sides show a varously distinct keel, that looks sharper at the left. Aperture deltoid, varying in outline and width. Body, extended - 60.26 ± 3.62 mm, less pigmented than in glabratus. Renal tube - 30.68 ± 1.69 mm, showing neither ridge nor pigmented line along its ventral surface, this negative character affording a sure means of separation from glabratus. Ovotestis - 14.48 ± 1.93 mm. Ovisperm duct - 13.04 ± 1.60 mm, including the non-unwound seminal vesicle. The latter was 0.97 ± 0,21 mm in greatest width. Carrefour - Resembling that of glabratus. Sperm duct - 21.36 ± 1.53 mm. Prostate - Prostate duct 7.14 ± 0.74 mm, collecting a row of long diverticula numbering 19.6 ± 3.1 and more separate than in glabratus. Last diverticulum generally bifurcate or arborescent, the remaining ones arborescent. Vas deferens - 28.68 ± 1.38. Ratio vas deferens/vergic sac = 6.8±0.8. Verge - 3.08 ± 0.28 mm long, 0.11 ± 0.02 mm wide. Vergic sac - 3.07 ± 0.28 mm long, about 0.20 mm wide. Ratio vergic sac/preputium = 0.84 ± 0.12. Preputium - 3.69 ± 0.47 mm long, 0.85 ± 0.10 mm wide. Albumen gland - Resembling taht of glabratus. Oviduct - 16.26 ± 1.41 mm, swollen at the cephalic end. Uterus - 13.24 ± 1.19 mm. Vagina - 1.70 ± 0.22 mm, swolen at the caudal portion. Spermatheca - 2.78 ± 0.40 mm long, 0.86 ± 0.16 mm wide. Spermathecal duct 1.11 ± 0.20 mm. Radula - 125 to 168 horizontal rows of teeth (mean 153.9 ± 8.4). Radula formula 28-1-28 to 36-1-36 (mean 31.8 ± 1.9). Mode formula 31-1-31. The morphological characteristics of the renal region and shell, and the great body length in the same condition of shell diameter, distinguish A. nigricans from the most related species A. glabratus, giving support to considering it a good species from a txonomic or phenotypic standpoint (morphospecies).

Physa Marmorata Guilding, 1828 (Pulmonata: Physidae)

A description of Physa marmorata Guilding, 1828, based on material collected at its type-locality, the Caribbean island of Saint Vincent, is presented. The shell is thin, horn-colored, surface very glossy, diaphanous. Spire acute, elevated; protoconch distinct, rounded-conical, reddish-brown; five not shouldered, broadly convex whorls with subobsolete spiral lines and thin growth lines. Aperture elongated, 1.4-2.0 times as long as the remaining shell length, narrow obovate-lunate; upper half acute-angled,lower half oval,narrowly rounded at the base, outer lip sharp, inner lip completely closing the umbilical region; a very distinct callus on the parietal wall; columellar lip with a low ridge gradually merging into the callus. ratios: shell width/shell length = 0.44 - 0.52 (mean 0.47); spire length /shell lenght = 0.33-0.41 (mean 0.39); aperture length/shell lenght = 0.59-0.67 (mean 0.62). Oral lappets laterally mucronate, foot spatulate with deeply pigmented acuminate tail. Mantle reflection with 6-10 short triangular dentations covering nearly half the right surface of the body whorl, and 4-6 covering a part of the ventral wall. Body surface with tiny dots of greenish-yellow pigment besides melanin. Renal tube tightly folded in toa zigzag course. Ovotestis diverticula acinous, laterally pressed against each other around a collecting canal. Ovispermiduct with well-developed seminal vesicle. oviduct highly convoluted, merging into a less convoluted nidamental gland which narrows to a funnel-shaped uterus and a short vagina. Spermathecal body oblong, more or less constricted in the middle and somewhat curved; spermathecal duct uniformly narrow, a little longer than be body. About 20 prostatic diverticula, simple, bifurcate or divided into a few short branches, distalmost ones assembled into a cluster. Penis long, nearly uniformly narrow; penial canal with lateral opening about the junction of its middle and lower thirds. Penial sheath with a bulbous terminal expasion the tip of which isinserted into the caudal end of the prepuce. Prepuce shouldered, much wider than the narrow portion of the penial sheath. Penial sheath/prepuce ratio about 2.08 (1.45-2.75). The main extrinsic muscles of the penial complex are a retractor, with a branch attached to the bulb, and another to the caudal end of the penial sheath; and a protractor, with a branch attached to the shoulder of the prepuce and adjoining area of the penial sheath, and another to the caudal end of the penial sheath. Egg capsule C-shaped, with 10-30 elliptical eggs (snails 10mm long) measuring about 1.10 mm (0.90-1.32) through the long axis and surrounded by an inner and an outer lamellate membranes. Jaw a simple obtusely V-shaped plate. radula will be described separately.

Biomphalaria obstructa (Morelet, 1849): a study of topotypic specimens (Mollusca: pulmonata: planorbidae)

A description of Biomphalaria obstructa (Morelet, 1849), based on specimens collected at its type locality - isla del carmen, state of Campeche, Mexico - is presented. The Shell is small, 13 mm in diameter, 3.5 mm in width and with 5.75 whorls in the largest specimen, thin, moderately lustrous and translucent, horn-colored. Whorls increasing regularly (neither slowly nor rapidly) in diameter, rounded on the periphery side, bluntly angular on the left. Suture well-marked, deeper on the left. Right side widely concave, with first whorl deeply situated and partly hidden by the next. Left side shallower than right one, largely flattened, with first whorl plaintly visible. Aperture roundly heart-shaped, usually in the same plane as the body whorl but somewhat deflected to the left (less frequently to the right) in some specimens. Peristome sharp, seldom blunt; a distinct callus on the parietal wall. A number of young shells develop one set (seldom more) of apertural lamellae which tend to be resorbed as the shell grows. Absence of renal ridge. Ovotestis with about 70 mostly unbrached diverticula. Seminal vesicle beset with well-developed knoblike to fingerlike diverticula. Vaginal pouch more or less developed. Spermatheca club-shaped when empty, egg-shaped when full, and with intermediate forms between those extremes. Spermathecal body usually somewhat longer than the duct. Prostate with 7 to 20 (mean 12.06 ± 2.51) usually short diverticula which give off plumpish branches spreading out in a fan shape and overlapping to some extent their immediate neighbors. Foremost prostatic diverticulum nearly always partially or completely inserted between the spermathecal body and the uterine wall. Penial sheath consistently narrower and shorter than the prepuce. Muscular coat of the penis consisting of an inner longitudinal and an outer circular layers. Ratios between organ lengths: caudal to cephalic parts of female duct = 0.55 to 1.37 (mean 0.85 +- 0.17); cephalic parte of female duct to penial complex = 1.36 to 2.81 ((mean 1.90 +- 0.33); penial sheath to prepuce = 042 to 0.96 (mean 0.67 +- 0.13). Comparison with Morelet’s type specimens of Planorbis orbiculus and P. retusus points to the identity of those nominal species with B. obstructa.

Possible hybridization of Brazilian planorbid snails and its importance in population dynamics

This study focuses on the possibility of experimental hybridization among host snail species for Schistosoma mansoni in Brazil, with morphological characterization of the hybrids found. By using albinism as a genetic marker, intraspecific crossbreedings were performed between two strains of each species involved, in addition to interspecific crossbreedings; the only viable crossbreeding was between pigmented Biomphalaria glabrata (Paulista, PE) and albino B. tenagophila (Joinville, SC), with the formation of F1 and F2 generations. All offspring in F1 displayed black eyes and a renal ridge on the mantle, while F2 displayed dissociated morphological traits. With regard to reproduction, F1 was more efficient than F2. The experiment's results suggest post-zygotic reproductive isolation.

Ootaxonomic investigation of five Lutzomyia species (Diptera, Psychodidae) from Venezuela

The eggshell fine structure of five sand fly species from Venezuela belonging to the genus Lutzomyia (L. migonei, L. ovallesi, L. absonodonta, L. gomezi and L. panamensis) was examined by scanning electron microscopy. The chorionic sculpturing of L. migonei, L. ovallesi, L. absonodonta and L. gomezi was characterized by series of columns arranged in palisade to form sinuous ridges. In inter-ridge areas, the basal layer was covered with fibrous material. The outer chorion of L. panamensis had a pattern known as "mountain- or volcano-like". The morphology of the posterior pole and aeropyle had a common structure in the five species, with some species-specific characters. The eggshell features of the five species are compared with those of other phlebotomine sand flies.

Phlebotomines (Diptera: Psychodidae) in forested areas of the Serra da Bodoquena, state of Mato Grosso do Sul, Brazil

Investigation was undertaken on the behaviour of the phlebotomine fauna in caves, forests, and anthropic environments of the Serra da Bodoquena, between January 1998 and January 2000. This paper reports on the phlebotomines captured in forested areas with automatic light traps (ALT), Shannon traps (ST), aspiration (AN), at natural resting sites and by human attractiveness (HA) during 24 h. The diversity and abundance of the species were investigated with ALT installed at 16 points (ground level) and 6 in the canopy. Natural infection by flagellates was investigated in females captured with ST, AN, and HA. The sand fly fauna was represented by 23 species. Twenty-two of these were captured with ALT, 15 of them on the western side, and 20 on the eastern. Lutzomyia longipalpis and Nyssomyia whitmani were the most abundant on the former and this species together with Lutzomyia almerioi on the latter side. On the eastern side the ecotopes located close to caves rendered a significantly greater number (P < 0.01) of specimens than did more distant sites. On this side Lu. almerioi contributed with 56% of the total number of specimens. Lu. almerioi females were predominantly attracted by humans (96.4%) and by ST (93.2%) and three of the 2173 dissected (0.138%) presented natural infection by flagellates. The attraction of Lu. almerioi to humans occurred during all seasons, predominantly in the summer, and in nocturnal and diurnal periods. Thus it is bothersome to inhabitants of and visitors to the Bodoquena ridge and a potential vector of flagellates.

Parasitism of two zoonotic reservoirs Dasyprocta leporina and D. fuliginosa (Rodentia) from Amazonas, with Trichostrongylina nematodes (Heligmonellidae): description of a new genus and a new species

A new genus and a new species of Heligmonellidae nematodes are described parasiting the stomach of three agoutis (two Dasyprocta fuliginosa and one D. leporina) captured in the middle and high Negro river microregion, state of Amazonas, Brazil. The new genus, as well as its type-species, are closely related to the trichostrongylids included in Fuellebornema, particularly on what concerns the pattern of the caudal bursa, but differing from them by the characteristics of the synlophe, that presents a poorly developed carene, when compared to the referred number of body ridges in Freitastrongylus n. gen. and consequently in F. angelae n. sp.,in which the ridges are well developed and the carene at mid-body has a similar size when compared to the ridge situated in front of the right field (ridge no. 5). Caudal bursa is of the type 1-4, with rays 9 shorter than rays 10, with a very long genital cone.

Spinitectus osorioi (Nematoda: Cystidicolidae) in the Mexican endemic fish Atherinella alvarezi (Atherinopsidae) from the Atlantic River drainage system in Chiapas, Southern Mexico

Specimens of Spinitectus osorioi Choudhury and Pérez-Ponce de León, an intestinal nematode species previously considered to be specific to Chirostoma spp and endemic to some lakes in the Pacific drainage in Michoacán, were collected from the freshwater fish Atherinella alvarezi (Díaz-Pardo) (Atherinopsidae) of the Michol River near Palenque, Chiapas, Southern Mexico, which belongs to the Atlantic drainage system. Studies using light and scanning electron microscopy revealed some taxonomically important, previously unreported or erroneously reported features of S. osorioi, such as the location of the vulva, the actual number and distribution of postanal papillae and phasmids and the presence of a short median cuticular ridge anterior to the cloacal opening (in addition to two long subventral ridges). The recorded somewhat shorter spicules (420-465 and 105-111 μm) and mostly smaller eggs (33-36 × 18-20 μm) as compared to the original species description may be due to a different type of host, geographical region or generally smaller body measurements of these specimens. These biometrical differences are considered to be within the limits of the intraspecific variability of S. osorioi. A key to species of Spinitectus parasitizing freshwater fishes in Mexico is provided.

Myxidium volitans sp. nov., a parasite of the gallbladder of the fish, Dactylopterus volitans (Teleostei: Triglidae) from the Brazilian Atlantic coast: morphology and pathology

Myxidium volitans sp. nov. (Myxozoa: Myxidiidae) parasitizing the hypertrophied green-brownish gallbladder of the teleost Dactylopterus volitans, collected in the Atlantic coast near Niterói, Brazil was described based on ultrastructural studies. The spores were fusiform, sometimes slightly crescent-shaped on average 21.7 ± 0.3 µm (mean ± standard deviation) (n = 50) long and 5.6 ± 0.4 µm (n = 30) wide. The spore wall was thin and smooth, comprising two equally-sized valves joined by a hardly visible sutural ridge. Spores containing two pyriform polar capsules (PC) (5.0 ± 0.4 × 2.3 ± 0.3 µm) (n = 30) are situated in each extremity of the spore. The PC wall was composed of hyaline layer (0.20-0.29 µm thick) and by a thin external granular layer. Each PC contains a polar filament (PF) with irregular arrangements that was projected from its apical region to the bases of PC and coiled laterally from bases to the tip of PC. Some regular striations and S-like structures in the periphery of the PFs with four-five irregular sections were observed. Based on the spore morphology, ultrastructural differences and the specificity of the host we describe this parasite as a new myxosporidian, named M. volitans sp. nov.

Phlebotomines (Diptera, Psychodidae) in caves of the Serra da Bodoquena, Mato Grosso do Sul State, Brazil

The present paper deals with the phlebotomine species captured during the period from January 1998 to June 2000 in 12 caves located in the Serra da Bodoquena, situated in the south central region of Mato Grosso do Sul State, Brazil. Three of the caves are situated further north (in Bodoquena county), seven in the central area (Bonito county) and two in the south (Jardim county). These last two caves and three of those in Bonito are located at the west side of the ridge. Eighteen species of phlebotomines were captured within the caves: Brumptomyia avellari (Costa Lima, 1932), Brumptomyia brumpti (Larrousse, 1920), Brumptomyia cunhai (Mangabeira, 1942), Brumptomyia galindoi (Fairchild & Hertig, 1947), Evandromyia corumbaensis (Galati, Nunes, Oshiro & Rego, 1989), Lutzomyia almerioi Galati & Nunes, 1999, Lutzomyia longipalpis (Lutz & Neiva, 1912), Martinsmyia oliveirai (Martins, Falcão & Silva, 1970), Micropygomyia acanthopharynx (Martins, Falcão & Silva, 1962), Micropygomyia peresi (Mangabeira, 1942), Micropygomyia quinquefer (Dyar, 1929), Nyssomyia whitmani (Antunes & Coutinho, 1939), Psathyromyia campograndensis (Oliveira, Andrade-Filho, Falcão & Brazil, 2001), Psathyromyia punctigeniculata (Floch & Abonnenc, 1944), Psathyromyia shannoni (Dyar, 1929), Pintomyia kuscheli (Le Pont, Martinez, Torrez-Espejo & Dujardin, 1998), Sciopemyia sordellii (Shannon & Del Ponte, 1927) and Sciopemyia sp. A total of 29,599 phlebotomine sandflies was obtained. Lutzomyia almerioi was absolutely predominant (91.5%) over the other species on both sides of the Bodoquena ridge, with the exception of the southern caves in which it was absent. It presents summer predominance, with nocturnal and diurnal activities. The species breeds in the caves and was captured during daytime both in the dark area and in the mouth of the caves. Martinsmyia oliveirai, the second most frequent sandfly, also presents a summer peak and only predominated over the other species in one cave, in which there were human residues.0