Efeitos de Dietas com Diferentes Níveis de Fibra em Detergente Neutro e do Tamanho de Poros de Sacos de Náilon Incubados no Rúmen sobre a Fauna Ruminal em Bubalinos e Bovinos

Quatro bubalinos e quatro bovinos, fistulados no rúmen, foram distribuídos aleatoriamente a dois quadrados latinos (4 x 4) para avaliar os efeitos de rações com quatro níveis de FDN (54, 60, 66 e 72%) e de dois tamanhos de poros em sacos de náilon (53 e 100 mm) sobre a população de protozoários ciliados no conteúdo ruminal e dentro dos sacos incubados em diversos tempos. Não houve diferença nos diversos gêneros estudados e no total de protozoários nas amostras dos sacos de náilon entre 53 e 100 mm. As concentrações de Entodinium e de total de protozoários do conteúdo ruminal elevaram-se até o nível de 66% de FDN, com queda acentuada em 72% nos bubalinos, enquanto nos bovinos houve redução com o aumento de FDN na ração. Houve diferenças nas concentrações dos protozoários da subfamília Diplodiniinae, Epidinium e Dasytricha entre bubalinos e bovinos, dentro das amostras incubadas em sacos com poros de 53 mm. A concentração de Diplodiniinae aumentou com 72% de FDN na dieta em ambas as espécies animais. Os bubalinos apresentaram valores médios de pH e taxa de passagem do líquido ruminal mais elevados e volumes ruminais menores que os bovinos.

Efeitos de dietas com polpa cítrica em substituição ao milho em grãos no concentrado sobre a degradabilidade e a fauna ruminal em bubalinos

Quatro búfalos com cânulas ruminais foram distribuídos aleatoriamente em quadrado latino 4 x 4, compreendendo quatro rações com feno de capim-coastcross (Cynodon dactylon) (50% MS) e níveis crescentes de polpa cítrica peletizada em substituição ao milho em grão moído no concentrado (0, 33, 66 e 100% MS). Foram avaliadas a degradabilidade dos nutrientes e a população de protozoários no rúmen. Houve efeito quadrático da fração solúvel dos nutrientes entre os tratamentos, com valores maiores nos dois extremos. A taxa de degradação da PB do feno e da MS da polpa cítrica apresentou-se linear com aumento da polpa cítrica na ração. As rações contendo polpa cítrica promoveram menores concentrações de Entodinium e número total de ciliados/mL que a dieta só com milho. Houve predominância dos Entodinium nas dietas contendo somente milho ou somente polpa cítrica, enquanto em mistura predominaram os ciliados da subfamília Diplodiniinae. Observou-se efeito de interação da polpa cítrica com o milho, promovendo modificações no metabolismo ruminal, especialmente na solubilidade dos nutrientes, na curva do pH e na composição da fauna.

DETECTION OF INFECTIOUS-DISEASE AGENTS IN TISSUE BY IMMUNOCYTOCHEMISTRY

1. Immunocytochemical procedures have played an increasingly larger role in the identification of infectious disease agents in tissue sections owing to the increased availability and specificity of antibody reagents, the great sensitivity of the methods, and the relative facility with which the studies are performed.2. Immunocytochemical methods can be applied to routine formalin-fixed tissue for the detection of infectious agents such as viruses, bacteria, fungi, and protozoa among other microorganisms for diagnostic and research purposes.

INTRAGASTRIC INFECTION OF CONVENTIONAL AND GERM-FREE MICE WITH GIARDIA-LAMBLIA

The effects of experimental infection with Giardia lamblia were studied in 30-day old conventional and germfree CFW mice (7 animals in each group) of both sexes. Cysts were observed in the feces of both groups 6 to 7 days after intragastric infection of each animal with about 2.5 x 10(5) G. lamblia trophozoites. Fecal cyst level was statistically higher in germfree mice (about 10(5) cysts/g feces) when compared with the conventional group (about 10(4) cysts/g feces). The peak of infection in the conventional group apparently occurred on the 10th day after infection as indicated by an increase of fecal weight and by histopathological examination. Intense infiltration of the lamina propria and high reactional hyperplasia of the lymphoid component were observed in the conventional group. There was no infiltration or hyperplasia in germfree infected mice and fecal weight was relatively constant throughout the experiment. These results suggest that, as is the case for other intestinal pathogenic protozoa, the intestinal microflora is indispensable for the expression of the pathogenicity but not for the multiplication of G. lamblia.

Antibodies to Neospora caninum and Toxoplasma gondii in domestic cats from Brazil

The occurrence of antibodies to Neospora caninum and Toxoplasmagondii was determined in 400 domestic cats (Siamese, Persian, and undetermined breeds) from the Municipality of Aracatuba, São Paulo, Brazil, through the indirect fluorescence antibody test (IFAT). of the 400 cats, 100 were seropositive to T. gondii (25%, titer >= 64) and 98 to N. caninum (24.5%, titer >= 16). The rate of seropositive cats for T. gondii was correlated with age (chi(2)=35.7; p < 0.001), with a higher number of infected animals at older ages. of the 219 cats younger than 1-year-old, 13.2% were seropositive for T. gondii, while 39.2% were positive in the 181 older animals. The presence of N. caninum was also correlated with age (divided by(2)=8.8; p < 0.01), with 18.7% (41/219) and 31.5% (57/181) of positive animals at ages below and above 12-month, respectively. An association between the occurrences of both protozoa in the felines was also observed (chi(2)=19.6; p < 0.001).

Detection of cross infections by Leishmania spp. and Trypanosoma spp. in dogs using indirect immunoenzyme assay, indirect fluorescent antibody test and polymerase chain reaction

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Parasitos e/ou comensais intestinais em pacientes neoplásicos submetidos à quimioterapia

The objective of this study was to verify the prevalence of intestinal parasites and/or commensals in the neoplastic patients undergoing chemotherapy. Stool samples were analyzed by the method of Lutz (1919) and Rugai (1954), in triplicate. This work was composed of three groups, the first one (GI) formed by neoplastic patients that are not undergoing chemotherapy, the second (GII) comprised patients who were undergoing chemotherapy, and the third group (GIII) consisting of patients who completed chemotherapy. A total of 30 patients (GI-5, GII-18 and GIII-7) were screened at the Assis Regional Hospital of the Unified Health System of Assis, São Paulo. Additional information on antiparasitic treatment and tumor type were obtained by questionnaire. The positivity was 66.7% (20 cases) for intestinal parasites and/or commensals. The helminths were Ascaris lumbricoides (36.7%), Hookworms (20%) and Hymenolepis diminuta (3.3%). Among the highlights are protozoan Giardia lamblia (46.7%), Entamoeba coli (6.7%), E. histolytica/dispar (3.3%), Endolimax nana (3.3%) and Iodameba butschlii (3.3%). The high frequency of intestinal parasites and/or commensals in the neoplastic patients can be attributed to poor personal hygiene and lack of immunity to reinfection and poor knowledge of the prophylaxis of infection by protozoa and helminths. The results indicate the necessity of adopting a new criterion for neoplastic patients undergoing chemotherapy, primarily performing parasitological diagnosis, treatment and monitoring of cure of intestinal parasitic infections in this risk group.

Ocorrência de enteroparasitos em centros de educação infantil no Município de Patos de Minas, MG, Brasil

Considering the high prevalence of the parasitic diseases in Brazil and its close relationship with the socio-economic and cultural levels of the population, aimed to establish the occurrence of enteroparasites in children of six Centers of Education Child (CEIs) of the municipality of Patos of Minas (MG). Were collected three samples of faeces of 161 children from zero to six years of age. Were used the Lutz and Baermann-Moraes' methods, to the detection of cysts of protozoa, eggs and larvae of helminths. Coproparasitological analyses showed positivity of 73%. The most frequent helminths were: Ascaris lumbricoides (50%), hookworm (22%) and Enterobius vermicularis (0.6%). Among the protozoa highlighted were: Entamoeba histolytica/E. dispar (22%) and Giardia lamblia (32%). The high occurrence of enteroparasites evidenced the need for an effective health policy in the CEIs of the municipality of Patos of Minas to the fight against intestinal parasites.

Parasitoses em lactentes de 0-12 meses atendidos na Faculdade de Medicina de Botucatu - São Paulo

Infestation by protozoa and/or helminths is considered to be extremely rare in infants. We therefore reviewed the records of all infants up to 12 mth of age attending the Paediatric Department from January 1973 to June 1977, in order to assess the frequency of stool examinations for parasites within this age group, the percentage of positive results in these examinations, and some personal characteristics which propitiated parasitic infestation. Some characteristics of the investigated infants were compared to those of a representative sample of the total number of infants attending during the same period. Stool examinations were realized in 11.8% of the 1,162 attending patients and of those, 15.3% were positive. The proportion of positive cases became 24.0% when the infants in whom there was a history of parasites being passed were included. Ascaris lumbricoides and Giardia lamblia were the parasites most frequently found, respectively in 42.8% and 37.1% of the positive cases. The only detected difference between the control group and the group of infants who had parasitologic examinations was the presence of diarrhea, significantly more frequent in the investigated group. The proportion of positive cases was significantly greater in girls, in infants older than 6 mth and when diarrhea was present for up to 15 days, than in boys, in infants up to 6 mth of age and in chronic diarrhea, respectively. Our results show that parasites were investigated in only a small percentage of infants under 1 yr of age, but that in this age group already, parasitic infestation does occur with a certain frequency. As investigated cases were selected, mainly due to the presence of diarrhea, the real prevalence of parasitism in infants under 1 yr of age was impossible to evaluate.

Targeting Plasmodium ligands on mosquito salivary glands and midgut with a phage display peptide library

Despite vast efforts and expenditures in the past few decades, malaria continues to kill millions of persons every year, and new approaches for disease control are urgently needed. To complete its life cycle in the mosquito, Plasmodium, the causative agent of malaria, has to traverse the epithelia of the midgut and salivary glands. Although strong circumstantial evidence indicates that parasite interactions with the two organs are specific, hardly any information is available about the interacting molecules. By use of a phage display library, we identified a 12-aa peptide-salivary gland and midgut peptide 1 (SM1)-that binds to the distal lobes of the salivary gland and to the luminal side of the midgut epithelium, but not to the midgut surface facing the hemolymph or to ovaries. The coincidence of the tissues with which parasites and the SM1 peptide interact suggested that the parasite and peptide recognize the same surface ligand. In support of this hypothesis, the SM1 peptide strongly inhibited Plasmodium invasion of salivary gland and midgut epithelia. These experiments suggest a new strategy for the genetic manipulation of mosquito vectorial capacity.

Comparação entre técnicas coproparasitológicas no diagnóstico de ovos de helmintos e oocistos de protozoários em cães.

In this study we evaluated the frequency of enteroparasites in pet dogs and their association with age, sex and breed, as well as the efficiency of the Willis-Mollay, Faust, Sedimentation and Direct exam methods. By these methods we processed 401 fecal samples. The samples were positive in the following percentages: Ancylostoma spp. (53.1%), Toxocara canis (20.7%), Cystoisospora ohioensis (15.7%), Trichuris vulpis (3.7%), Dipylidium caninum (2.5%) and Taenia spp. (1.0%). Toxocara canis (67.3%) and C. ohioensis (47.3%) showed higher positivity in the puppies. The Willis-Mollay technique was more efficient in the diagnosis of Ancylostoma spp. and T. canis eggs. The Direct method was the least efficient. It was found that the majority of the cases of D. caninum were diagnosed by the Sedimentation method (8=2.0%), while for T. vulpis Willis-Mollay (12=3.0%) and Sedimentation (13=3.2%) were more efficient. In view of these results, we can recommend the association of Willis-Mollay and Sedimentation methods for the diagnosis of gastrointestinal helminths. Due to the elevated occurrence of Ancylostoma spp. and T. canis, which are involved in zoonotic diseases, it becomes necessary to apply more efficient prophylaxis of canine intestinal parasitosis at the City of Araçatuba, state of São Paulo.

Serological profile of anti-Toxoplasma gondii antibodies in apparently healthy dogs of the city of Botucatu, São Paulo state, Brazil

Toxoplasmosis is a zoonosis caused by Toxoplasma gondii, a coccidian protozoan of worldwide distribution. The seroprevalence in canine population can be an alternative for measuring T. gondii urban spreading. A total of 780 blood samples from dogs were collected, during the yearly anti-rabies campaign, carried out by the Department of Veterinary Hygiene and Public Health, School of Veterinary Medicine and Animal Husbandry (FMVZ), São Paulo State University, UNESP, together with the county health authorities, in August 1999. Using Indirect Fluorescent Antibody Test (IFAT) for detecting antibodies anti-T. gondii in the sera samples, we observed that 258 dogs (33.1%) were positive. The associations between the serological results and the epidemiological variables were studied. Statistically significant differences were not found regarding sex (32.2% male and 34.3% female reactors). Dogs without a defined breed showed seropositivity statistically higher than the pedigreed group. The occurrence of infection was considered higher with age.

Toxoplasma gondii: Detection by mouse bioassay, histopathology, and polymerase chain reaction in tissues from experimentally infected pigs

In the present study, we evaluated three techniques, mouse bioassay, histopathology, and polymerase chain reaction (PCR) to detect Toxoplasma gondii infection in tissues from experimentally infected pigs. Twelve mixed breed pigs, seronegative for T. gondii using an indirect immunofluorescent antibody test (IFAT), were used. Ten pigs were infected with 4 × 104 VEG strain oocysts, and two were maintained as uninfected controls. Animals were killed 60 days pos infection. Muscle (heart, tongue, diaphragm, and masseter) and brain samples were collected to investigate the presence of T. gondii tissue cysts by the different assay methods. For the bioassay, samples of brain (50 g) and pool of muscle samples (12.5 g of tongue, masseter, diaphragm, and heart) were used. PCR was performed using Tox4 and Tox5 primers which amplified a 529 bp fragment. The DNA extraction and PCR were performed three times, and all tissue samples were tested individually (brain, tongue, masseter, diaphragm, and heart). For histopathology, fragments of tissues were fixed in 10% of buffered formal saline and stained with HE. Histopathological results were all negative. PCR showed 25/150 (16.6%) positive samples, being 17/120 (14.1%) and 8/30 (26.6%) from muscle, and brain tissues, respectively. Tissue cysts of T. gondii were identified by mouse bioassay in 54/98 (55.1%) samples, being 31/48 (64.6%) from muscle samples, and 23/50 (46.0%) from brain samples. Toxoplasma gondii isolation in muscle samples by mouse bioassay was higher than in PCR (P < 0.01). Results indicate that DNA from pig tissues interfered with 529-bp-PCR sensitivity, and mouse bioassay was better than PCR in detecting T. gondii in tissues from pigs. © 2006 Elsevier Inc. All rights reserved.