Um surto de coccidiose em perdizes (Rhynchotus rufescens), criadas em cativeiro, por Eimeria rhynchoti Reis e Nóbrega, 1936 (Apicomplexa: Eimeriidae).

Eimeria rhynchoti is redescribed parasitizing partridge (Rhynchotus rufescens), reared in captivity, from Jaboticabal City, São Paulo State, Brazil. Sporulation takes place in 48 hours, the shape of oocysts found vary from spherical to elliptic with 23.01 micro +/- 1.57 of length by 21.0 micro +/- 1.78 of width. The microple, polar cap and residuum of the oocysts were absent. The oocyst wall, measures 2.2 micro +/- 0.31 of thickness, is composed by two smooth layers; the polar granule is present. The sporocysts length was 15.03 mm +/- 2.12 by 8.08 mm +/- 0.84 of width vary from elliptic to elongate. Sporocyst wall slender with is fine and Stieda body; the residue found in form of several smaller granules spherical compacts. The sporozoites are contrary extending along the sporocysts wall possessing refracts body of easy visualization.

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.

Avaliação do kit TF-Test para o diagnóstico das infecções por parasitas gastrintestinais em ovinos

This study was performed to standardize parasite egg counting in feces of sheep by TF-Test, in addition to compare this test to the Gordon & Whitlock technique (G&W). Twenty-four lambs were artificially infected with Haemonchus contortus throughout 12 weeks. At the end of this time, faecal samples were taken and animals were slaughtered for worm identification and counting. G&W and TF-Test methods were carried out on each fecal sample. Both tests showed Haemonchus eggs in 95.8% of the samples (P>0.05). The correlation coefficients (r) between fecal egg counts (FEC) using G&W × Total Worm Count (TWC) were r=0.52 (not transformed data) and r=0.85 (transformed data); between FEC by TF-Test × TWC were r=0.51 (not transformed data) and r=0.87 (transformed data). Other 100 fecal samples were taken from naturally infected sheep. In these animals, the G&W and TF-Test methods showed 85% and 86% of fecal samples positive for Strongylidea eggs, respectively (P>0.05). Also in those animals, Eimeria oocysts were found in 33% of fecal samples by TF-Test, whereas in the G&W only 12% were positive (P<0.001). For Strongyloides spp., TF-Test showed 15% of positive fecal samples, whereas G&W showed 5% (P<0.05). In conclusion, both methods were efficient to diagnose gastrointestinal nematodes and TF-Test was superior to diagnose oocysts of Eimeria spp. and eggs of Strongyloides spp; conversely, Strongylidea eggs counting using TF-Test was underestimated.

Toxoplasma gondii in semen of experimentally infected swine

Eight reproductive boars were divided into three groups and inoculated with Toxoplasma gondii [GI (n=3) 1.5×104 oocysts strain P; GII (n=3) 1.0×106 tachyzoites strain RH; and GIII (n=2) non-inoculated control]. Clinical, hematological, parasitemia and serological tests and studies of the parasite in the semen through bioassay and PCR, and in reproductive organs (Bioassay and immunohistochemical analyses) were conducted to evaluate the toxoplasmic infection. Blood and semen were collected on day -2, -1, 1, 3, 5, 7, 9, 11, 14 and weekly up to 84 days post-inoculation (DPI). No clinical or hematimetric alteration was observed in the boars. Parasitemia was detected in one boar inoculated with oocysts at the 7th DPI and in another boar infected with tachyzoites (GII) at the 3rd and 49 th DPI. Serological tests revealed antibodies against T. gondii in animals inoculated with oocysts or tachyzoites at the 7th DPI with dilutions of 1:256 and 1:64, which reached peaks of 1:4096 at day 11 and 9, respectively. The bioassays revealed the presence of the parasite in semen samples of a boar inoculated with oocysts (GI) at 3, 49 and 56 DPI and from two boars infected with tachyzoites (GII), one animal at 5 and two animals at 49 days DPI. Mice inoculated with semen from the control group (GIII) remained serologically negative. PCR analysis showed T. gondii DNA in the semen of Boar 1 and Boar 3 inoculated with tachyzoites and oocysts, respectively. The immunohistochemical tests showed T. gondii in the reproductive organs of Boar 1 and Boar 2, inoculated with tachyzoites and oocysts, respectively. These findings suggest the possible occurrence of venereal transmission of T. gondii in swine.

Cryptosporidium SP in HIV-infected individuals attending a Brazilian university hospital

The aim of the current work was to evaluate the occurrence of Cryptosporidium sp in AIDS patients in a region of São Paulo State, Brazil. Patients were divided into groups according to CD4+ T lymphocyte count and use of potent antiretroviral treatment. Two hundred and ten fecal samples from 105 patients were fixed in 10% formalin and subjected to centrifuge formol-ether sedimentation. Slides were stained with auramine and confirmed by modified Ziehl-Neelsen. Cryptosporidiosis occurrence was 10.5% with no relationship among gender, age or the presence of diarrhea. The number of oocysts in all samples was small, independent of CD4+ T lymphocyte count, HIV plasma viral load, and presence of diarrhea. These results may be due to the reduced prevalence of opportunistic infections in AIDS individuals after the advent of highly active antiretroviral therapy.

Evaluation of IFA, MAT, ELISAs and immunoblotting for the detection of anti-Toxoplasma gondii antibodies in paired serum and aqueous humour samples from experimentally infected pigs

The study evaluated the efficiency of diagnostic laboratory methods to detect anti-Toxoplasma gondii antibodies in paired serum and aqueous humour samples from experimentally infected pigs. 18-mixed breed pigs were used during the experiment; these were divided into two groups, G1 (infected group, n = 10) and G2 (uninfected group, n = 8). Infection was performed with 4 × 10 4 VEG strain oocysts at day 0 by the oral route in G1 animals. All pigs were euthanized at day 60, when retina, aqueous humour, and blood samples were collected. Anti-T. gondii antibody levels were assessed in serum (s) and aqueous humour (ah) by indirect immunofluorescence assay (IFA), modified agglutination test (MAT), m-ELISA (using crude membranes from T. gondii tachyzoites as antigen) and r-ELISA (using rhoptries from T. gondii tachyzoites as antigen). Polymerase chain reactions (PCR) of samples from the retina were performed by using Tox4 and Tox5 primers. Antibody titers of G1 animals ranged from 128 to 1024 and from 16 to 256 in serum and aqueous humour, respectively. There were differences in the correlation coefficients between IFA(s) × IFA (ah) (r = 0.62, P = 0.05), MAT(s) × MAT (ah) (r = 0.97, P < 0.0001); however, there was no significant difference between r-ELISA(s) × r-ELISA (ah) (r = 0.14, P = 0.7). Antibodies present in serum and aqueous humour recognized similar antigens. Samples of retina were positive by PCR in 30% (3/10) of infected pigs. G2 animals remained without antibody levels and were PCR negative throughout the experiment. These results suggest that the use of a combination of tests and immunoblotting for paired aqueous humour and serum samples could improve the sensitivity and specificity for the diagnosis of ocular toxoplasmosis. © 2007 Elsevier Ltd. All rights reserved.

Ocorrência de parasitas gastrintestinais em cães e gatos na rotina do laboratório de enfermidades parasitárias da FMVZ/Unesp-Botucatu, SP.

With the object of determining the occurrence and prevalence of parasites in dogs and cats in the region of Botucatu, a survey was conducted from the results of fecal parasitologic exams feces, processed by the laboratory of parasitic diseases of FMVZ-UNESP/Botucatu from January 2002 to December 2006. 1,012 fecal samples of dogs and cats were evaluated by the technique of Willis Mollay and Faust. In dogs the higher incidence was found for Ancylostoma caninum eggs (38%). In cats, the oocysts of Isospora spp were present in 48.38% of positive samples.

Ações de vigilância continuada, papel do cão como animal sentinela para toxoplasmose.

Toxoplasmosis is a worldwide zoonosis caused by Toxoplasma gondii and its definitive host is the domestic and wild felids infecting human beings and other warmblooded animals. Dogs are considered a potential risk on the transmission due they can mechanically transmit oocysts to man. In this study, a retrospective analysis of toxoplasmic infection in dog serum samples sent to Serviço de Diagnóstico de Zoonoses/FMVZ-UNESP/Botucatu, SP, in the period of 1998 to 2007 was performed. During this period 1097 serum samples were analyzed by the indirect fluorescent antibody test (IFAT), with 299 (27.25%) positive. The most frequent titer was 16 (42.80%), followed by 64 (37.79%). The results indicate that T.gondii is distributed in the environment showing the role of the dog as sentinel animal to toxoplasmosis to monitor public health actions to the control of this zoonosis.

Eimeria stiedai: Metabolism of lipids, proteins and glucose in experimentally infected rabbits, Oryctolagus cuniculus

Rabbits were experimentally infected with sporulated Eimeria stiedai oocysts. A total of 50 white adult rabbits, New Zealand race, were distributed into two groups: Group A was infected with 1x10 4 sporulated Eimeria stiedai oocysts, while group B was inoculated with distilled water as a control. The animals generally displayed increased levels of total protein, globulin, total cholesterol, LDL-c and triacylglycerols; however, total levels of liver lipids and HDL-c decreased, and plasma glucose levels varied during the experimental period. In sum, Eimeria stiedai infection of rabbits caused a considerable number of changes in the metabolism of lipids, proteins and glucose, which is likely due to direct effects of liver cirrhosis on normal body function.

Serological survey of Toxoplasma gondii in captive Neotropical felids from Southern Brazil

Toxoplasma gondii is the causative intracellular protozoan of toxoplasmosis in human being and animals. Members of the Felidae family are considered the single definitive host for the infection; both wild and domestic cats are able to excrete oocysts in the environment. Wild cats maintained in captivity may serve as source of infection for other clinically susceptible animals in the same environment. The aim of this study was to determine the frequency of T. gondii IgG antibodies in 57 neotropical felids (1 Leopardus geoffroyi; 3 Puma yagouaroundi; 17 Leopardus wiedii; 22 Leopardus tigrinus; and 14 Leopardus pardalis) kept at the Bela Vista Biological Sanctuary, Itaipu Binacional, Southern Brazil, by the modified agglutination test (MAT) using titer 16 as cut-off point. Seropositivity was observed in 38/57 (66.67%; 95% CI 53.66-77.51%) samples, with higher frequency in ocelots (71.43%). Wild-caught felids were three times more likely to be infected when compared to zoo-born animals (P≤ 0.05) and age of wild-caught animals (P= 0.6892; 95% CI. = 0.7528-1.66) was not significant as a risk factor for the infection, the same occurring with zoo-born animals (P= 0.05; 95% CI. = 0.6267-24.052). These results suggest that, despite efforts to control T. gondii infection in zoo facilities, such as individual pens, hygiene monitoring, veterinary care and pre-frozen meat offered as food, non-domestic felids kept in captivity, particularly the wild-caught specimens, may be invariably exposed to infection due to other environmental sources. © 2010 Elsevier B.V.

Identificação de espécies e genótipos de Cryptosporidium em bovinos leiteiros no Brasil

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

Importância da criptosporidiose como zoonose

The protozoa of the genus Cryptosporidium sp. are parasites that complete their life cycle on the surface of epithelial cells of the respiratory, gastrointestinal and urinary tracts of mammals, birds, reptiles and fish. Affected animals generally are asymptomatic but gastrointestinal disorders sometimes can be present. Criptoporidiosis is considered a zoonosis. Infected animals, especially cattle, are a source of infection for the environment and humans, because they eliminate large numbers of oocysts in their feces. The symptoms when present is characterized by watery diarrhea, dehydration, abdominal pain, weight loss and death, especially in immunosuppressed individuals, and especially in patients with human immunodeficiency virus (HIV) and children. Therapeutic methods effective to eliminate this agent in animals and humans has not been developed but it is necessary to applied support treatment. Criptosporidiosis is considered by the World Health Organization as an emerging disease. Basic sanitation, use of appropriate methods for the inactivation of oocysts and security of personal hygiene are recommended as Prophylactic methods to minimize the spread of Cryptosporidium. This literature review was aimed at describing the importance of cryptosporidiosis in public health.

Coccidiose em cães e gatos do município de andradina, estado de são paulo, Brasil

Coccidiosis is one of the most common parasitic diseases in dogs and cats in all the world. The aim of this study was to determine the occurrence of this parasitosis in dog and cat population at the Municipality of Andradina in the State of São Paulo, from 2007 to 2009. Fecal samples from 97 cats and 93 dogs were analyzed by using the techniques of flotation in saturated sodium chloride and spontaneous sedimentation. The species were classified according to morphology of the oocysts. Cystoisospora fecal oocyst found in 71.1% (69/97) of the cats, and simple infection by C. rivolta and C. felis occurred respectively in 41.0% (16/39) and 20.5% (8/39) animals, with P ≥ 0.2319. In 39.7%(37/93) of the dogs was found positive for Cystoisospora spp. And the species C. canis identified in the largest proportion (63.9%) with P = 0.0005. From the results, we conclude that dogs and cats had high incidence of infection Cystoisospora, being C. canis and C. rivolta most observed species, respectively.

Parasitism level by helminths and weight gain of calves kept in organic and conventional grazing

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Sexual transmission of Toxoplasma gondii in sheep

Male sheep of reproductive age were distributed into three groups: GI, a sheep inoculated (oral) with 2.0×105 oocysts of the P strain of Toxoplasma gondii; GII, a sheep infected (subcutaneous) with 1.0×106 tachyzoites of the RH strain of T. gondii; and GIII, a sheep kept as a control (not infected). After the inoculation of the males, 12 breeding ewes, which were not pregnant and which were serologically negative for reproductive diseases (particularly toxoplasmosis), were distributed into three groups, synchronized, and subsequently exposed to natural mating with previously inoculated males. The distribution was as follows: five ewes that underwent natural mating with the GI male, five ewes that were exposed to natural mating with the GII male, and two ewes that were mated with the non-infected male (control). Serum samples of all the ewes were collected on days -30, -14, -7, -1, and 0 (days before natural mating) and on days 1, 3, 5, 7, 11, 14, and weekly until birth; the presence of serum antibodies against T. gondii was assessed by IFAT. Using a bioassay and PCR, T. gondii was isolated from the semen of the infected reproducing sheep before mating. Following natural mating, 5 of the 12 females displayed antibodies specific for T. gondii; of these animals, two of the ewes underwent natural mating with the male inoculated with oocysts (GI) and three with the male infected with tachyzoites (GII). One of the females that displayed antibodies specific to this coccidian and that underwent natural mating with the GII sheep had a macerated fetus on the 70th day following coverage. Using a bioassay after the birth, it was possible to isolate T. gondii from samples of the pool of tissues from the five females that seroconverted after natural mating and from their respective lambs. Using PCR, the DNA of T. gondii was isolated from the pool of tissues from one and two females exposed to natural mating with the reproductive males infected with the oocysts and tachyzoites, respectively. Using this technique, it was also possible to diagnose the presence of the parasite in the pool of tissues from the lambs of one female that underwent natural mating with the male sheep infected with oocysts. These results demonstrated the sexual transmission of T. gondii in the sheep species with consequent vertical transmission to their lambs. © 2013 Elsevier B.V.