Sôbre u'a modificação do meio de Monteverde

It is well known that the culture media used in the presumptive diagnosis of suspiciuous colonies from plates inoculated with stools for isolation of enteric organisms do not always correctly indicate the major groups of enterobacteria. In an effort to obtain a medium affording more exact indications, several media (1-9) have been tested. Modifications of some of these media have also been tested with the result that a satisfactory modification of Monteverde's medium was finaly selected. This proved to be most satisfactory, affording, as a result of only one inoculation, a complete series of basic indications. The modification involves changes in the formula, in the method of preparation and in the manner of storage. The formulae are: A. Thymol blue indicator: NaOH 0.1/N .............. 34.4 ml; Thymol blue .............. 1.6 g; Water .................... 65.6 ml. B. Andrade's indicator. C. Urea and sugar solution: Urea ..................... 20 g; Lactose ................... 30 g; Sucrose ................... 30 g; Water .................... 100 ml. The mixture (C.) should be warmed slightly in order to dissolve the ingredients rapidly. Sterilise by filtration (Seitz). Keep stock in refrigeratior. The modification of Monteverde's medium is prepared in two parts. Semi-solid part - Peptone (Difco) 2.0 g; NaCl 0.5 g; Agar 0.5 g; Water 100.0 ml. Boil to dissolve the ingredients. Adjust pH with NaOH to 7.3-7.4. Boil again for precipitation. Filter through cotton. Ad indicators "A" 0.3 ml and "B" 1.0 ml. Sterilise in autoclave 115ºC, 15 minutes in amounts not higher than 200 ml. Just before using, add solution "C" asseptically in amounts of 10 ml to 200 ml of the melted semi-solid medium, maintained at 48-50ºC. Solid part - Peptone (Difco) 1.5 g; Trypticase (BBL) 0.5 g; Agar 2.0 g; Water 100,00 ml. Boil to dissolve the ingredients. Adjust pH with NaOH to 7.3-7.4. Boils again. Filter through cotton. Add indicators "A" 0.3 ml and "B" 1.0 ml; ferrous ammonium sulfate 0.02 g; sodiun thiosulfate 0.02 g. Sterilise in autoclave 115ºC, 15 minutes in amounts not higher than 200 ml. Just before using, add solution "C" asseptically in amounts of 10 ml to 200 ml of the melted solid medium, maintained at 48-50ºC. Final medium - The semi-solid part is dispensed first (tubes about 12 x 120 mm) in 2.5 ml amounts and left to harden at room temperature, in vertical position. The solid part is dispensed over the hardened semi-solid one in amounts from 2.0 ml to 2.5 ml and left to harden in slant position, affording a butt of 12 to 15 mm. The tubes of medium should be subjected to a sterility test in the incubator, overnight. Tubes showing spontaneous gas bubbles (air) should then be discarded. The medium should be stored in the incubator (37ºC), for not more than 2 to 4 days. Storage of the tubes in the ice-box produces the absorption of air which is released as bubbles when the tubes are incubated at 37ºC after inoculation. This fact confirmed the observation of ARCHAMBAULT & McCRADY (10) who worked with liquid media and the aplication of their observation was found to be essential to the proper working conditions of this double-layer medium. Inoculation - The inoculation is made by means of a long straight needle, as is usually done on the triple sugar, but the needel should penetrate only to about half of the height of the semi-solid column. Indol detection - After inoculation, a strip of sterelized filter papaer previously moistened with Ehrlich's reagent, is suspended above the surface of the medium, being held between the cotton plug and the tube. Indications given - In addition to providing a mass of organisms on the slant for serological invetigations, the medium gives the following indications: 1. Acid from lactose and/or sucrose (red, of yellowsh with strains which reduce the indicators). 2. Gas from lactose and/or sucrose (bubbles). 3. H[2]S production, observed on the solid part (black). 4. Motility observed on the semi-solid part (tubidity). 5. Urease production, observed on solid and semi-solid parts (blue). 6. Indol production, observed on the strip of filter paper (red or purplish). Indol production is not observed with indol positive strains which rapidly acidify the surface o the slant, and the use of oxalic acid has proved to give less sensitive reaction (11). Reading of results - In most cases overnight incubation is enough; sometimes the reactions appear within only a few hours of incubation, affording a definitive orientation of the diagnosis. With some cultures it is necessary to observe the medium during 48 hours of incubation. A description showing typical differential reaction follows: Salmonella: Color of the medium unchanged, with blackening of the solid part when H[2]S is positive. The slant tends to alkalinity (greenish of bluish). Gas always absent. Indol negative. Motility positive or negative. Shigella: Color of the medium unchanged at the beginning of incubation period, but acquiring a red color when the strain is late lactose/sucrose positive. Slant tending to alkalinity (greenish or purplish). Indol positive or negative. Motility, gas and H[2]S always negative. Proteus: Color of the medium generally changes entirely to blue or sometimes to green (urease positive delayed), with blackening of solid part when H[2]S is positive. Motility positive of negative. Indol positive. Gas positive or negative. The strains which attack rapidly sucrose may give a yellow-greenish color to the medium. Sometimes the intense blue color of the medium renders difficult the reading of the H[2]S production. Escherichiae and Klebsiellae: Color of the medium red or yellow (acid) with great and rapid production of gas. Motility positive or negative. Indol generally impossible to observe. Paracoli: Those lactose of sucrose positive give the same reaction as Esherichia. Those lactose or sucrose negatives give the same reactions as Salmonellae. Sometimes indol positive and H[2]S negative. Pseudomonas: Color of the medium unchanged. The slant tends to alkalinity. It is impossible to observe motility because there is no growth in the bottom. Alkaligenes: Color of the medium unchanged. The slant tends to alkalinity. The medium does not alter the antigenic properties of the strains and with the mass of organisms on the slant we can make the serologic diagnosis. It is admitted that this medium is somewhat more laborious to prepare than others used for similar purposes. Nevertheless it can give informations generally obtained by two or three other media. Its use represents much saving in time, labor and material, and we suggest it for routine laboratory work in which a quick presumptive preliminary grouping of enteric organisms is needed.

Corythaica cyathicollis (Costa, 1864), aspectos sistemáticos, biológicos e econômicos (Hemiptera, Tingidae)

The present work deals with the systematic, biological and economic problems related to Corythaica cyathicollis (Costa, 1864) (Hemip., Tingidae). In the first part are presented the generic characteristics of Corythaica and is discussed the status of the specific name. The validity of C. cyathicollis, as stated by DRAKE and his collaborators, was denied by MONTE in his last works, he considered the species as C. passiflorae. Even in the modern literature no agreement has been achieved and three names are still used (cyathicollis, passiflorae and planaris) to designate the same insect. In order to resolve definitively this problem, a Neotype is designed to fill the place of the missing type of C. cyathicollis. Also in the first parte is discussed the taxonomic value of both male and female genitalia. The whole male copulator apparatus is studied and are illustrated the genital capsules of 8 species of this genus. Special mention is made of the shape of the basal plates and the proportions of the segmental membrana. The female genitalia is studied based upon the work of FELDMAN & BAILEY (1952). In the second part the biological cycle of C. cyathicollis is carefully studied. Descriptions of the egg are done and the ways of oviposition. The number of eggs laid by the female was observed to be about 350, during a period of more than 45 days. The eclosion of the neanide I is illustrated in some of its phases and the 5 larval instars are described and illustrated. Ending this part are included the lists of parasites and predators observed as well as the plant hosts. The actual geographical distribution is presented, based chiefly on HURD (1945). The economic problems concerning this species are reported in the third part of the work, and the ways of control are discussed. An experiment was carried out involving 4 insecticides: Malathion and Parathion, commonly used against this "lace bug"; Toxaphene and Dimethoate (American Cyanamid 12.880), the last one is an insecticide recently introduced in Brazil and was not previously used for these purposes, but gave the best results and it is quite able to control these insects even on crops showing highly developed infestations.

Sôbre o Phlebotomus Brasiliensis Costa Lima, 1932 (Diptera, Psychodidae)

In 1939, Mangabeira obtained, under laboratory conditions, the development of eggs of Phlebotomus brasiliensis Costa Lima, 1932, collected at Lassance (typical locality), Minas Gerais, Brasil. He then studied the female and immature stages of this Phlebotomus. The results of these observations plus some more recent data on the male, geographical distribution and bionomics are presented. Morphologically it is closest to Phlebotomus runoides. However, the male Phlebotomus brasiliensis differs from all other Phlebotomus because of its very long spicules, similar to those of Brumptomyia. The female differs by its longer ducts, and by possessing only four horizontal teeth in the buccal cavity, whereas P. runoides has approximately 12 teeth. The pupae of P. brasiliensis is characterized by its two pre-alar setae, which are very simple and small and by the abdominal setae, which are not planted on a protruding tubercle. The fourth stage larvae main characteristics are very thin antennae, inserted on a protruding tuberculum, and slightly brush-like hind frontal setae. P. brasiliensis is here reported, for the first time, for the State of Bahia (Cachoeira, Pojuca and Salvador). The species has almost always been found in armadillo burrows. In the State of Bahia it is more frequent during the dry season. Under laboratory conditions, the female lays about 53 eggs.

Sôbre dois novos parasitos de peixe da costa do Peru (Cestoda, Tetraphyllidea)

Na presente nota estudamos alguns cestódeos coletados de Sarda chilensis (Cuv.) peixe comum da costa do pacífico, enviado por um de nós (N.I.H); o material era constituído por vários espécimes de Phyllobothriidae e paenas trêss de Onchobothriidae, infelizmente todos comprimidos; não obstante aqui fazemos o seu estudo por ser tratar de material de interêsse.

Revisão do gênero Klossinemella Costa, 1961 (Nematoda, Cobboldinidae), com descrição de uma nova subfamília e de uma nova espécie

Neste trabalho os autores discutem a posição sistemática do gênero Klossinemella Costa, 1961, propondo uma nova organização para a família Cobboldinidae Skrjabin, 1948. Apresentam a descrição das espécies dêste gênero (K. iheringi, K. conciliatus e K. travassosi sp. n.) e do ciclo evolutivo.

Trematódeos monogenéticos (Polistomatas) da costa brasileira I. sobre Loimos scitulus sp. n. (Loimoidae) e Tagia ecuadori (Meserve, 1938) Sproston, 1946 (Diclidophoridae)

Em continuação aos estudos dos trematódeos monogenéticos da Coleção Helmintológica do Instituto Oswaldo Cruz, descrevemos no presente trabalho uma espécie do gênero Loimos MacCallum, 1917, Loimolinae Price, 1936, loimoidae Bychowsky, 1957 que consideramos nova para a ciência, e assinalamos nova ocorrência de Tagia ecuadori (Meserve, 1938) Sproston, 1946, Tagiinae Yamaguti, 1963, Diclidophoridae Najibina & Obonikova, 1971, no Atlântico Sul. Loimos scitulus sp. n. diferencia-se das outras espécies do gênero pelos seguintes caracteres: forma e estrututra do proaptor, oótipo grande, número de testículos, posição do poro genital, filamento do ovo e forma de opistaptor. Dentre as diferenças dadas Loimos scitulus sp. n. aproxima-se de L. salpinggoides pela estrutura do proaptor, de L. secundus pela posição do poro genital; de L. winteri pelo opistaptor. Quanto aos exemplares de Tagia ecuadori por nós estudados, apesar de ter sido evidenciada uma vagina, identificamos a esta espécie, por apresentar estruturas e medidas que se enquadram nas variações dadas pelos estudiosos do grupo.